Journal: Molecular cell

Article Title: CRISPR-Cas13 inhibitors block RNA-editing in bacteria and mammalian cells

doi: 10.1016/j.molcel.2020.03.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides crRNAs This study Table S4 Recombinant DNA Plasmids This study Table S5 Antibodies mouse monoclonal GFP antibody Biolegend Cat# 668206 anti-his tag anti-body Thermo Fisher Scientific Cat# TA150087 Bacterial and Virus Strains E. coli c-3000 ATCC Cat# 15597 NEB 5-alpha Competent E. coli New England BioLabs Cat# C2987H BL21 Competent E. coli New England BioLabs Cat# C2530H E. coli bacteriophage MS2 ATCC Cat# 15597-B1 Leptotrichia buccalis DSM 1135 ATCC Cat# 14201 E. coli c3000-derived strains This study Table S6 Chemicals, Peptides, and Recombinant Proteins LB Broth Base Thermo Fisher Scientific Cat# 12780029 LB Agar Fisher Scientific Cat# BP1425-500 SOC medium New England BioLabs Cat# B9020S ampicillin Fisher Scientific Cat#11593027 chloramphenicol Sigma Cat# C0378 kanamycin Fisher Scientific Cat# 25389-94-0 gentamicin ACROS Cat# 1405-41-0 IPTG Invitrogen Cat# 15529019 penicillin-streptomycin Thermo Fisher Scientific Cat# 10378016 fetal bovine serum VWR Seradigm Cat# 97068-085 LipofectAmine 3000 Thermo Fisher Scientific Cat# L3000015 Opti-MEM I Reduced Serum Medium Thermo Fisher Scientific Cat# 11058021 RMPI 1640 Medium Thermo Fisher Scientific Cat# 11875119 protease inhibitor cockt Thermo Fisher Scientific Cat# 78425 Critical Commercial Assays QIAGEN Plasmid Mini Kit QIAGEN Cat# 12125 myTXTL Sigma 70 Master Mix Kit Arbor Bioscience Cat# 507024 AMPure XP beads Beckman Coulter Cat# A63882 DynabeadsTM Sheep anti-Mouse IgG Invitrogen Cat# 11201D PierceTM His Protein Interaction Pull-Down Kit Thermo Fisher Scientific Cat# 21277 PierceTM Gaussia-Firefly Luciferase Dual Assay Kit Thermo Fisher Scientific Cat# 16181 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 Maxima SYBR Green qPCR Master Mix Thermo Fisher Scientific Cat# K0252 Direct-zolTM RNA MiniPrep kit Zymo Reseach Cat# R2052 the CellTiter 96 AQueous One Solution Cell Proliferation assay Promega Cat# G3580 Pierce LDH Cytotoxicity Assay Kit Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C20300","term_id":"1632571","term_text":"C20300"}} C20300 Experimental Models: Cell Lines HEK293T ATCC Cat# CRL-3216 Open in a separate window KEY RESOURCES TABLE An integrated approach is designed to screen anti-CRISPRs (AcrVIA) candidates A series of prophages are discovered in regions encoding AcrVIAs AcrVIAs inhibit Cas13a RNA targeting in bacteria and human cells AcrVIAs impede programmable RNA base editing by blocking dCas13a-based toolkit

Techniques: Recombinant, Virus, Protease Inhibitor, Plasmid Preparation, Luciferase, Reverse Transcription, SYBR Green Assay, Proliferation Assay, LDH Cytotoxicity Assay

Journal: Molecular cell

Article Title: CRISPR-Cas13 inhibitors block RNA-editing in bacteria and mammalian cells

doi: 10.1016/j.molcel.2020.03.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides crRNAs This study Table S4 Recombinant DNA Plasmids This study Table S5 Antibodies mouse monoclonal GFP antibody Biolegend Cat# 668206 anti-his tag anti-body Thermo Fisher Scientific Cat# TA150087 Bacterial and Virus Strains E. coli c-3000 ATCC Cat# 15597 NEB 5-alpha Competent E. coli New England BioLabs Cat# C2987H BL21 Competent E. coli New England BioLabs Cat# C2530H E. coli bacteriophage MS2 ATCC Cat# 15597-B1 Leptotrichia buccalis DSM 1135 ATCC Cat# 14201 E. coli c3000-derived strains This study Table S6 Chemicals, Peptides, and Recombinant Proteins LB Broth Base Thermo Fisher Scientific Cat# 12780029 LB Agar Fisher Scientific Cat# BP1425-500 SOC medium New England BioLabs Cat# B9020S ampicillin Fisher Scientific Cat#11593027 chloramphenicol Sigma Cat# C0378 kanamycin Fisher Scientific Cat# 25389-94-0 gentamicin ACROS Cat# 1405-41-0 IPTG Invitrogen Cat# 15529019 penicillin-streptomycin Thermo Fisher Scientific Cat# 10378016 fetal bovine serum VWR Seradigm Cat# 97068-085 LipofectAmine 3000 Thermo Fisher Scientific Cat# L3000015 Opti-MEM I Reduced Serum Medium Thermo Fisher Scientific Cat# 11058021 RMPI 1640 Medium Thermo Fisher Scientific Cat# 11875119 protease inhibitor cockt Thermo Fisher Scientific Cat# 78425 Critical Commercial Assays QIAGEN Plasmid Mini Kit QIAGEN Cat# 12125 myTXTL Sigma 70 Master Mix Kit Arbor Bioscience Cat# 507024 AMPure XP beads Beckman Coulter Cat# A63882 Dynabeads™ Sheep anti-Mouse IgG Invitrogen Cat# 11201D Pierce™ His Protein Interaction Pull-Down Kit Thermo Fisher Scientific Cat# 21277 Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit Thermo Fisher Scientific Cat# 16181 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 Maxima SYBR Green qPCR Master Mix Thermo Fisher Scientific Cat# K0252 Direct-zol™ RNA MiniPrep kit Zymo Reseach Cat# R2052 the CellTiter 96 AQueous One Solution Cell Proliferation assay Promega Cat# G3580 Pierce LDH Cytotoxicity Assay Kit Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C20300","term_id":"1632571","term_text":"C20300"}} C20300 Experimental Models: Cell Lines HEK293T ATCC Cat# CRL-3216 Open in a separate window KEY RESOURCES TABLE An integrated approach is designed to screen anti-CRISPRs (AcrVIA) candidates A series of prophages are discovered in regions encoding AcrVIAs AcrVIAs inhibit Cas13a RNA targeting in bacteria and human cells AcrVIAs impede programmable RNA base editing by blocking dCas13a-based toolkit

Techniques: Recombinant, Virus, Protease Inhibitor, Plasmid Preparation, Luciferase, Reverse Transcription, SYBR Green Assay, Proliferation Assay, LDH Cytotoxicity Assay

Journal: Molecular cell

Article Title: CRISPR-Cas13 inhibitors block RNA-editing in bacteria and mammalian cells

doi: 10.1016/j.molcel.2020.03.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides crRNAs This study Table S4 Recombinant DNA Plasmids This study Table S5 Antibodies mouse monoclonal GFP antibody Biolegend Cat# 668206 anti-his tag anti-body Thermo Fisher Scientific Cat# TA150087 Bacterial and Virus Strains E. coli c-3000 ATCC Cat# 15597 NEB 5-alpha Competent E. coli New England BioLabs Cat# C2987H BL21 Competent E. coli New England BioLabs Cat# C2530H E. coli bacteriophage MS2 ATCC Cat# 15597-B1 Leptotrichia buccalis DSM 1135 ATCC Cat# 14201 E. coli c3000-derived strains This study Table S6 Chemicals, Peptides, and Recombinant Proteins LB Broth Base Thermo Fisher Scientific Cat# 12780029 LB Agar Fisher Scientific Cat# BP1425-500 SOC medium New England BioLabs Cat# B9020S ampicillin Fisher Scientific Cat#11593027 chloramphenicol Sigma Cat# C0378 kanamycin Fisher Scientific Cat# 25389-94-0 gentamicin ACROS Cat# 1405-41-0 IPTG Invitrogen Cat# 15529019 penicillin-streptomycin Thermo Fisher Scientific Cat# 10378016 fetal bovine serum VWR Seradigm Cat# 97068-085 LipofectAmine 3000 Thermo Fisher Scientific Cat# L3000015 Opti-MEM I Reduced Serum Medium Thermo Fisher Scientific Cat# 11058021 RMPI 1640 Medium Thermo Fisher Scientific Cat# 11875119 protease inhibitor cockt Thermo Fisher Scientific Cat# 78425 Critical Commercial Assays QIAGEN Plasmid Mini Kit QIAGEN Cat# 12125 myTXTL Sigma 70 Master Mix Kit Arbor Bioscience Cat# 507024 AMPure XP beads Beckman Coulter Cat# A63882 Dynabeads™ Sheep anti-Mouse IgG Invitrogen Cat# 11201D Pierce™ His Protein Interaction Pull-Down Kit Thermo Fisher Scientific Cat# 21277 Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit Thermo Fisher Scientific Cat# 16181 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 Maxima SYBR Green qPCR Master Mix Thermo Fisher Scientific Cat# K0252 Direct-zol™ RNA MiniPrep kit Zymo Reseach Cat# R2052 the CellTiter 96 AQueous One Solution Cell Proliferation assay Promega Cat# G3580 Pierce LDH Cytotoxicity Assay Kit Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C20300","term_id":"1632571","term_text":"C20300"}} C20300 Experimental Models: Cell Lines HEK293T ATCC Cat# CRL-3216 Open in a separate window KEY RESOURCES TABLE An integrated approach is designed to screen anti-CRISPRs (AcrVIA) candidates A series of prophages are discovered in regions encoding AcrVIAs AcrVIAs inhibit Cas13a RNA targeting in bacteria and human cells AcrVIAs impede programmable RNA base editing by blocking dCas13a-based toolkit

Techniques: Recombinant, Virus, Protease Inhibitor, Plasmid Preparation, Luciferase, Reverse Transcription, SYBR Green Assay, Proliferation Assay, LDH Cytotoxicity Assay

Journal: Molecular cell

Article Title: CRISPR-Cas13 inhibitors block RNA-editing in bacteria and mammalian cells

doi: 10.1016/j.molcel.2020.03.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides crRNAs This study Table S4 Recombinant DNA Plasmids This study Table S5 Antibodies mouse monoclonal GFP antibody Biolegend Cat# 668206 anti-his tag anti-body Thermo Fisher Scientific Cat# TA150087 Bacterial and Virus Strains E. coli c-3000 ATCC Cat# 15597 NEB 5-alpha Competent E. coli New England BioLabs Cat# C2987H BL21 Competent E. coli New England BioLabs Cat# C2530H E. coli bacteriophage MS2 ATCC Cat# 15597-B1 Leptotrichia buccalis DSM 1135 ATCC Cat# 14201 E. coli c3000-derived strains This study Table S6 Chemicals, Peptides, and Recombinant Proteins LB Broth Base Thermo Fisher Scientific Cat# 12780029 LB Agar Fisher Scientific Cat# BP1425-500 SOC medium New England BioLabs Cat# B9020S ampicillin Fisher Scientific Cat#11593027 chloramphenicol Sigma Cat# C0378 kanamycin Fisher Scientific Cat# 25389-94-0 gentamicin ACROS Cat# 1405-41-0 IPTG Invitrogen Cat# 15529019 penicillin-streptomycin Thermo Fisher Scientific Cat# 10378016 fetal bovine serum VWR Seradigm Cat# 97068-085 LipofectAmine 3000 Thermo Fisher Scientific Cat# L3000015 Opti-MEM I Reduced Serum Medium Thermo Fisher Scientific Cat# 11058021 RMPI 1640 Medium Thermo Fisher Scientific Cat# 11875119 protease inhibitor cockt Thermo Fisher Scientific Cat# 78425 Critical Commercial Assays QIAGEN Plasmid Mini Kit QIAGEN Cat# 12125 myTXTL Sigma 70 Master Mix Kit Arbor Bioscience Cat# 507024 AMPure XP beads Beckman Coulter Cat# A63882 Dynabeads™ Sheep anti-Mouse IgG Invitrogen Cat# 11201D Pierce™ His Protein Interaction Pull-Down Kit Thermo Fisher Scientific Cat# 21277 Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit Thermo Fisher Scientific Cat# 16181 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 Maxima SYBR Green qPCR Master Mix Thermo Fisher Scientific Cat# K0252 Direct-zol™ RNA MiniPrep kit Zymo Reseach Cat# R2052 the CellTiter 96 AQueous One Solution Cell Proliferation assay Promega Cat# G3580 Pierce LDH Cytotoxicity Assay Kit Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C20300","term_id":"1632571","term_text":"C20300"}} C20300 Experimental Models: Cell Lines HEK293T ATCC Cat# CRL-3216 Open in a separate window KEY RESOURCES TABLE An integrated approach is designed to screen anti-CRISPRs (AcrVIA) candidates A series of prophages are discovered in regions encoding AcrVIAs AcrVIAs inhibit Cas13a RNA targeting in bacteria and human cells AcrVIAs impede programmable RNA base editing by blocking dCas13a-based toolkit

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Luciferase, SYBR Green Assay, Proliferation Assay, LDH Cytotoxicity Assay

Journal: Molecular cell

Article Title: CRISPR-Cas13 inhibitors block RNA-editing in bacteria and mammalian cells

doi: 10.1016/j.molcel.2020.03.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides crRNAs This study Table S4 Recombinant DNA Plasmids This study Table S5 Antibodies mouse monoclonal GFP antibody Biolegend Cat# 668206 anti-his tag anti-body Thermo Fisher Scientific Cat# TA150087 Bacterial and Virus Strains E. coli c-3000 ATCC Cat# 15597 NEB 5-alpha Competent E. coli New England BioLabs Cat# C2987H BL21 Competent E. coli New England BioLabs Cat# C2530H E. coli bacteriophage MS2 ATCC Cat# 15597-B1 Leptotrichia buccalis DSM 1135 ATCC Cat# 14201 E. coli c3000-derived strains This study Table S6 Chemicals, Peptides, and Recombinant Proteins LB Broth Base Thermo Fisher Scientific Cat# 12780029 LB Agar Fisher Scientific Cat# BP1425-500 SOC medium New England BioLabs Cat# B9020S ampicillin Fisher Scientific Cat#11593027 chloramphenicol Sigma Cat# C0378 kanamycin Fisher Scientific Cat# 25389-94-0 gentamicin ACROS Cat# 1405-41-0 IPTG Invitrogen Cat# 15529019 penicillin-streptomycin Thermo Fisher Scientific Cat# 10378016 fetal bovine serum VWR Seradigm Cat# 97068-085 LipofectAmine 3000 Thermo Fisher Scientific Cat# L3000015 Opti-MEM I Reduced Serum Medium Thermo Fisher Scientific Cat# 11058021 RMPI 1640 Medium Thermo Fisher Scientific Cat# 11875119 protease inhibitor cockt Thermo Fisher Scientific Cat# 78425 Critical Commercial Assays QIAGEN Plasmid Mini Kit QIAGEN Cat# 12125 myTXTL Sigma 70 Master Mix Kit Arbor Bioscience Cat# 507024 AMPure XP beads Beckman Coulter Cat# A63882 Dynabeads™ Sheep anti-Mouse IgG Invitrogen Cat# 11201D Pierce™ His Protein Interaction Pull-Down Kit Thermo Fisher Scientific Cat# 21277 Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit Thermo Fisher Scientific Cat# 16181 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 Maxima SYBR Green qPCR Master Mix Thermo Fisher Scientific Cat# K0252 Direct-zol™ RNA MiniPrep kit Zymo Reseach Cat# R2052 the CellTiter 96 AQueous One Solution Cell Proliferation assay Promega Cat# G3580 Pierce LDH Cytotoxicity Assay Kit Thermo Fisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C20300","term_id":"1632571","term_text":"C20300"}} C20300 Experimental Models: Cell Lines HEK293T ATCC Cat# CRL-3216 Open in a separate window KEY RESOURCES TABLE An integrated approach is designed to screen anti-CRISPRs (AcrVIA) candidates A series of prophages are discovered in regions encoding AcrVIAs AcrVIAs inhibit Cas13a RNA targeting in bacteria and human cells AcrVIAs impede programmable RNA base editing by blocking dCas13a-based toolkit

Techniques: Recombinant, Virus, Protease Inhibitor, Plasmid Preparation, Luciferase, Reverse Transcription, SYBR Green Assay, Proliferation Assay, LDH Cytotoxicity Assay