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ad cre gfp adenovirus  (Vector Biolabs)


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    Vector Biolabs ad cre gfp adenovirus
    Ad Cre Gfp Adenovirus, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ad+cre+gfp+adenovirus/Ad-Cre-GFP/pmc12465051-263-10-12
    Average 96 stars, based on 162 article reviews
    ad cre gfp adenovirus - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Infection:

    Article Title: LIN28-mediated gene regulatory loops synchronize developmental transitions throughout organogenesis
    Article Snippet: .. Lin28a/b were knocked out post-neurosphere formation via adenoviral infection with Ad-Cre-GFP adenovirus (Vector Biolabs, 1700) or Ad-GFP adenovirus (Vector Biolabs, 1060). ..

    Article Title: LIN28-mediated gene regulatory loops synchronize transitions throughout organogenesis
    Article Snippet: .. Lin28a/b were knocked out post-neurosphere formation via adenoviral infection with Ad-Cre-GFP adenovirus (Vector Biolabs, 1700) or Ad-GFP adenovirus (Vector Biolabs, 1060). ..

    Article Title: Neratinib, a pan ERBB/HER inhibitor, restores sensitivity of PTEN -null, BRAFV600E melanoma to BRAF/MEK inhibition
    Article Snippet: .. PTEN KO clones and the parental A375 line were then infected with Ad-Cre-GFP adenovirus (approximate MOI 30, resulting in ~95%–99% GFP positive cells) from Vector Biolabs (Cat. No. 1700). ..

    Article Title: Neratinib, a pan ERBB/HER inhibitor, restores sensitivity of PTEN -null, BRAFV600E melanoma to BRAF/MEK inhibition.
    Article Snippet: .. PTEN KO clones and the parental A375 line were then infected with Ad-Cre-GFP adenovirus (approximate MOI 30, resulting in ~95%–99% GFP positive cells) from Vector Biolabs (Cat. No. 1700). ..

    Purification:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    Virus:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    Subcloning:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    Recombinant:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    HAT Assay:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    SYBR Green Assay:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    Polymerase Chain Reaction:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    CRISPR:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    TA Cloning:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    cDNA Synthesis:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..


    Sequencing:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Western Blot:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: List of DNA oligos used for CRISPR-mediated gene editing, off-target analyses, and variant transcript sequencing. .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignalTM West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Plasmid Preparation:

    Article Title: POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status
    Article Snippet: Images were analyzed and percent confluency was measured using IncuCyte Software (Sartorius). .. ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti-mouse IgG Sigma-Aldrich Cat# F0257; RRID:AB_259378 Purified anti-BrdU Biolegend Cat# 317902; RRID:AB_604040 Phospho-p53 (Ser15) mouse Cell Signaling Technology Cat# 9286; RRID:AB_331741 Anti-p53 (DO-1) Santa Cruz Biotechnology Cat# SC-126; RRID:AB_628082 Anti-p21 Abcam Cat# CP74; RRID:AB_1603138 Anti-MDM2 (SMP14) Santa Cruz Biotechnology Cat# SC-965; RRID:AB_627920 Anti-PUMAα/β (H-136) Santa Cruz Biotechnology Cat# SC-28226; RRID:AB_2064827 Bacterial and Virus Strains Subcloning Efficiency DH5α Competent Cells Invitrogen Cat# 18265017 Ad-Cre-GFP Adenovirus, Cre Recombinase Vector Biolabs Cat# 1700 Biological Samples N/A Chemicals, Peptides, and Recombinant Proteins UltraPure Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) Invitrogen Cat# 15593031 KpnI-HF New England Biolabs Cat# R3142S SacI-HF New England Biolabs Cat# R3138S EcoRI-HF New England Biolabs Cat# R3101S NotI-HF New England Biolabs Cat# R3189S XbaI Invitrogen Cat# IVGN0126 T7 Endonuclease I New England Biolabs Cat# M0302S Lipofectamine 2000 Invitrogen Cat# 11668030 Geneticin Life Technologies Cat# 10131027 HAT Supplement (50X) Life Technologies Cat# 21060017 6-Thioguanine Sigma Cat# A4882-250MG Crystal Violet ThermoFisher Scientific Cat# C581-100 MEM Non-Essential Amino Acids Solution (100X) Life Technologies Cat# 11140050 0.05% Trypsin-EDTA Life Technologies Cat# 25300120 Trizol Reagent Invitrogen Cat# 15596018 DEPC-Treated Water Invitrogen Cat# AM9915G 5-Bromo-2′-deoxyuridine Sigma Cat# B5002 Propidium iodide Sigma Cat# P4170 SuperSignal™ West Pico PLUS Chemiluminescent Substrate ThermoFisher Scientific Cat# 34578 Critical Commercial Assays SsoAdvanced Universal SYBR Green Supermix Bio-Rad Cat# 1725270 PureLink PCR Purification Kit Invitrogen Cat# K310002 GeneArt CRISPR Nuclease Vector with OFP Reporter Kit Invitrogen Cat# A21174 TOPO TA Cloning Kit for Subcloning Invitrogen Cat# 451641 First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Cat# 27926101 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat# 5000006 Deposited Data Genomic Data Commons Data Portal National Cancer Institute GDC Data Portal https://portal.gdc.cancer.gov/ Whole Exome Sequencing Raw Datasets This paper SRA accession number: PRJNA589337 Unprocessed Western blot images Mendeley doi: 10.17632/cdgs6fmpsn.2 Experimental Models: Cell Lines Human: HCT116 Other RRID: CVCL_0291 Human: HCT116+Mlh1 This paper (also Hodel et al 2018 eLife) N/A Human: HCT116 POLE wt/S459F This paper N/A Human: HCT116 POLE wt/S459F +Mlh1 This paper N/A Human: HCT116 POLE S459F/S459F +Mlh1 This paper N/A Human: HCT116 POLE wt/P286R +Mlh1 This paper N/A Experimental Models: Organisms/Strains N/A Oligonucleotides POLE guide sequences, see Supp. ..

    Clone Assay:

    Article Title: Neratinib, a pan ERBB/HER inhibitor, restores sensitivity of PTEN -null, BRAFV600E melanoma to BRAF/MEK inhibition
    Article Snippet: .. PTEN KO clones and the parental A375 line were then infected with Ad-Cre-GFP adenovirus (approximate MOI 30, resulting in ~95%–99% GFP positive cells) from Vector Biolabs (Cat. No. 1700). ..

    Article Title: Neratinib, a pan ERBB/HER inhibitor, restores sensitivity of PTEN -null, BRAFV600E melanoma to BRAF/MEK inhibition.
    Article Snippet: .. PTEN KO clones and the parental A375 line were then infected with Ad-Cre-GFP adenovirus (approximate MOI 30, resulting in ~95%–99% GFP positive cells) from Vector Biolabs (Cat. No. 1700). ..



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    Cell proliferation in mouse bladder tumor organoids depends on the catalytic activity of KMT9. ( A ) Schematic of the establishment of bladder tumor organoids. Bladders of Pten fl/fl / Trp 53 fl/fl , Pten fl/fl / Trp 53 fl/fl / Kmt9α fl/fl , and Pten fl/fl / Trp53 fl/fl / Kmt9αN122A flKI/flKI mice were isolated, and single cells were grown under organoid culture conditions. Gene ablation was induced in vitro by infection of organoids with <t>adenovirus</t> driving Cre-GFP expression. GFP-positive cells isolated by Fluorescence-Activated Cell Sorting (FACS) were allowed to regrow as organoids. Organoids from C57BL/6 mice served as control. ( B ) Validation of depletion of PTEN, TRP53, and KMT9α by Western blot with the indicated antibodies. ( C ) Representative pictures ( left ) and relative size ( right ) of Ctrl, Pten / Trp53 KO, Pten / Trp53 / Kmt9α KO, and Pten / Trp53 KO/ Kmt9αN122A KI bladder (tumor) organoids 5 days after FACS. Scale bars, 100 μm. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant). ( D ) Relative cell viability of organoid cells grown as 2D cultures determined by MTT assay on day 4. Data represent means + s.d. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant), n = 3. ( E ) Pie chart showing the number of DEGs in Pten / Trp53 KO compared with Pten / Trp53 KO/ Kmt9αN122A KI organoids (8048) with numbers of upregulated and downregulated genes ( p < 1 × 10 4 ). ( F ) Enriched gene sets for GO biological processes obtained for the 8048 DEGs. ( G ) Heat maps displaying relative mRNA levels of genes involved in “regulation of cell cycle” (GO:0051726) ( left ) and “regulation of cell migration” (GO:0030334) ( right ) in Pten / Trp53 KO and Pten / Trp53 KO/ Kmt9αN122A KI cells. ( H ) Quantitative real-time PCR analysis of mRNA expression for selected genes presented in ( G ). Data represent means + standard deviation. Statistical significance was evaluated by a two-tailed t test, (*, p < 0.05; ***, p < 0.001) n = 3. ( B ) The original Western blots with molecular weight markers and original data of densitometry scans are presented in .
    Adenovirus, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    adenoviruses adeno cre gfp  (Vector Biolabs)
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    Vector Biolabs adenoviruses adeno cre gfp
    Cell proliferation in mouse bladder tumor organoids depends on the catalytic activity of KMT9. ( A ) Schematic of the establishment of bladder tumor organoids. Bladders of Pten fl/fl / Trp 53 fl/fl , Pten fl/fl / Trp 53 fl/fl / Kmt9α fl/fl , and Pten fl/fl / Trp53 fl/fl / Kmt9αN122A flKI/flKI mice were isolated, and single cells were grown under organoid culture conditions. Gene ablation was induced in vitro by infection of organoids with <t>adenovirus</t> driving Cre-GFP expression. GFP-positive cells isolated by Fluorescence-Activated Cell Sorting (FACS) were allowed to regrow as organoids. Organoids from C57BL/6 mice served as control. ( B ) Validation of depletion of PTEN, TRP53, and KMT9α by Western blot with the indicated antibodies. ( C ) Representative pictures ( left ) and relative size ( right ) of Ctrl, Pten / Trp53 KO, Pten / Trp53 / Kmt9α KO, and Pten / Trp53 KO/ Kmt9αN122A KI bladder (tumor) organoids 5 days after FACS. Scale bars, 100 μm. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant). ( D ) Relative cell viability of organoid cells grown as 2D cultures determined by MTT assay on day 4. Data represent means + s.d. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant), n = 3. ( E ) Pie chart showing the number of DEGs in Pten / Trp53 KO compared with Pten / Trp53 KO/ Kmt9αN122A KI organoids (8048) with numbers of upregulated and downregulated genes ( p < 1 × 10 4 ). ( F ) Enriched gene sets for GO biological processes obtained for the 8048 DEGs. ( G ) Heat maps displaying relative mRNA levels of genes involved in “regulation of cell cycle” (GO:0051726) ( left ) and “regulation of cell migration” (GO:0030334) ( right ) in Pten / Trp53 KO and Pten / Trp53 KO/ Kmt9αN122A KI cells. ( H ) Quantitative real-time PCR analysis of mRNA expression for selected genes presented in ( G ). Data represent means + standard deviation. Statistical significance was evaluated by a two-tailed t test, (*, p < 0.05; ***, p < 0.001) n = 3. ( B ) The original Western blots with molecular weight markers and original data of densitometry scans are presented in .
    Adenoviruses Adeno Cre Gfp, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell proliferation in mouse bladder tumor organoids depends on the catalytic activity of KMT9. ( A ) Schematic of the establishment of bladder tumor organoids. Bladders of Pten fl/fl / Trp 53 fl/fl , Pten fl/fl / Trp 53 fl/fl / Kmt9α fl/fl , and Pten fl/fl / Trp53 fl/fl / Kmt9αN122A flKI/flKI mice were isolated, and single cells were grown under organoid culture conditions. Gene ablation was induced in vitro by infection of organoids with adenovirus driving Cre-GFP expression. GFP-positive cells isolated by Fluorescence-Activated Cell Sorting (FACS) were allowed to regrow as organoids. Organoids from C57BL/6 mice served as control. ( B ) Validation of depletion of PTEN, TRP53, and KMT9α by Western blot with the indicated antibodies. ( C ) Representative pictures ( left ) and relative size ( right ) of Ctrl, Pten / Trp53 KO, Pten / Trp53 / Kmt9α KO, and Pten / Trp53 KO/ Kmt9αN122A KI bladder (tumor) organoids 5 days after FACS. Scale bars, 100 μm. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant). ( D ) Relative cell viability of organoid cells grown as 2D cultures determined by MTT assay on day 4. Data represent means + s.d. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant), n = 3. ( E ) Pie chart showing the number of DEGs in Pten / Trp53 KO compared with Pten / Trp53 KO/ Kmt9αN122A KI organoids (8048) with numbers of upregulated and downregulated genes ( p < 1 × 10 4 ). ( F ) Enriched gene sets for GO biological processes obtained for the 8048 DEGs. ( G ) Heat maps displaying relative mRNA levels of genes involved in “regulation of cell cycle” (GO:0051726) ( left ) and “regulation of cell migration” (GO:0030334) ( right ) in Pten / Trp53 KO and Pten / Trp53 KO/ Kmt9αN122A KI cells. ( H ) Quantitative real-time PCR analysis of mRNA expression for selected genes presented in ( G ). Data represent means + standard deviation. Statistical significance was evaluated by a two-tailed t test, (*, p < 0.05; ***, p < 0.001) n = 3. ( B ) The original Western blots with molecular weight markers and original data of densitometry scans are presented in .

    Journal: Cancers

    Article Title: Lysine Methyltransferase 9 (KMT9) Is an Actionable Target in Muscle-Invasive Bladder Cancer

    doi: 10.3390/cancers16081532

    Figure Lengend Snippet: Cell proliferation in mouse bladder tumor organoids depends on the catalytic activity of KMT9. ( A ) Schematic of the establishment of bladder tumor organoids. Bladders of Pten fl/fl / Trp 53 fl/fl , Pten fl/fl / Trp 53 fl/fl / Kmt9α fl/fl , and Pten fl/fl / Trp53 fl/fl / Kmt9αN122A flKI/flKI mice were isolated, and single cells were grown under organoid culture conditions. Gene ablation was induced in vitro by infection of organoids with adenovirus driving Cre-GFP expression. GFP-positive cells isolated by Fluorescence-Activated Cell Sorting (FACS) were allowed to regrow as organoids. Organoids from C57BL/6 mice served as control. ( B ) Validation of depletion of PTEN, TRP53, and KMT9α by Western blot with the indicated antibodies. ( C ) Representative pictures ( left ) and relative size ( right ) of Ctrl, Pten / Trp53 KO, Pten / Trp53 / Kmt9α KO, and Pten / Trp53 KO/ Kmt9αN122A KI bladder (tumor) organoids 5 days after FACS. Scale bars, 100 μm. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant). ( D ) Relative cell viability of organoid cells grown as 2D cultures determined by MTT assay on day 4. Data represent means + s.d. Statistical significance was evaluated by a one-way ANOVA test (***, p < 0.001; ns, not significant), n = 3. ( E ) Pie chart showing the number of DEGs in Pten / Trp53 KO compared with Pten / Trp53 KO/ Kmt9αN122A KI organoids (8048) with numbers of upregulated and downregulated genes ( p < 1 × 10 4 ). ( F ) Enriched gene sets for GO biological processes obtained for the 8048 DEGs. ( G ) Heat maps displaying relative mRNA levels of genes involved in “regulation of cell cycle” (GO:0051726) ( left ) and “regulation of cell migration” (GO:0030334) ( right ) in Pten / Trp53 KO and Pten / Trp53 KO/ Kmt9αN122A KI cells. ( H ) Quantitative real-time PCR analysis of mRNA expression for selected genes presented in ( G ). Data represent means + standard deviation. Statistical significance was evaluated by a two-tailed t test, (*, p < 0.05; ***, p < 0.001) n = 3. ( B ) The original Western blots with molecular weight markers and original data of densitometry scans are presented in .

    Article Snippet: For the in vitro deletion of the floxed alleles, organoids were treated with Cre-GFP-expressing adenovirus (1 × 10 10 pfu/mL, Vectorbiolabs, Malvern, PA, USA, #1700, Lot A0501T#11M18).

    Techniques: Activity Assay, Isolation, In Vitro, Infection, Expressing, Fluorescence, FACS, Control, Biomarker Discovery, Western Blot, MTT Assay, Migration, Real-time Polymerase Chain Reaction, Standard Deviation, Two Tailed Test, Molecular Weight