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lenti dcas9 krab blast  (Addgene inc)


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    Addgene inc lenti dcas9 krab blast
    Lenti Dcas9 Krab Blast, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lenti+dcas9+krab+blast/Lenti-dCas9-KRAB-blast+(Plasmid+%2389567)/pmc13657374-111-70-76
    Average 96 stars, based on 146 article reviews
    lenti dcas9 krab blast - by Bioz Stars, 2026-11
    96/100 stars

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    Related Articles

    CRISPR:

    Article Title: CRISPR screens identify PRMT7 as a therapeutic target to enhance T cell-mediated killing in breast cancer
    Article Snippet: .. For CRISPR screens and validations, breast cell lines were transduced with Lenti-Cas9-2A-Blast (Addgene, Cat# 73310) or Lenti-dCas9-KRAB-Blast (Addgene, Cat# 89567). .. MCF7 cells stably expressing Cas9 or KRAB-dCas9 were established by transduction of the virus at MOI < 0.7 , selected in 10 μg/ml blasticidin (Gibco, Cat# A1113903) and maintained in 5 μg/ml blasticidin.

    Article Title: CRISPR screens identify PRMT7 as a therapeutic target to enhance T cell-mediated killing in breast cancer.
    Article Snippet: .. Pooled CRISPR screens for resistance to T cell cytotoxicity For CRISPR screens and validations, breast cell lines were transduced with Lenti-Cas9-2A-Blast (Addgene, Cat# 73310) or Lenti-dCas9-KRAB-Blast (Addgene, Cat# 89567). .. MCF7 cells stably expressing Cas9 or KRAB-dCas9 were established by transduction of the virus at MOI<0.7 51, selected in 10 μg/ml blasticidin (Gibco, Cat# A1113903) and maintained in 5 μg/ml blasticidin.

    Article Title: BRRIAR lncRNA alters breast cancer risk by modulating interferon signaling in cis and in trans
    Article Snippet: Absorbance was measured at 450 nm using a Tecan i-control plate reader and concentrations calculated by interpolation using standard curves. .. Lenti-dCas9-KRAB-blast (a gift from Gary Hon [Addgene; #89567]), Lenti-Cas9‐2 A‐Blast plasmid (a gift from Jason Moffat [Addgene; #73310]), pgRNA-humanized (a gift from Stanley Qi [Addgene; #44248])or Human Brunello CRISPR knockout pooled library (a gift from David Root and John Doench [Addgene; #73178]) were co-transfected with two package plasmids, pMD2.G (a gift from Didier Trono [Addgene; #12259]) and psPAX2 (a gift from Didier Trono [Addgene; #12260]) into HEK293 cells using FuGENE6 (Promega). ..

    Article Title: HIV-1 transcription dominates over host gene activity at the HIV-1 integration site
    Article Snippet: .. For CRISPR-ready HIV-1-infected Jurkat T cell clones, HIV-1-infected Jurkat T cell clones (1G2, 8B10, 1D7, 1A8, 2F5, 5F9, and 1D9) were transduced with pseudotyped lentiviral vectors Lenti-MPHv2 (containing EF1α-driven MS2-p65-HSF1 activator and hygromycin selection marker, Addgene #89308) ( ) for CRISPRa and Lenti-dCas9-KRAB-blast (containing blasticidin selection marker, Addgene #89567) ( ) for CRISPRi, respectively. ..

    Transduction:

    Article Title: CRISPR screens identify PRMT7 as a therapeutic target to enhance T cell-mediated killing in breast cancer
    Article Snippet: .. For CRISPR screens and validations, breast cell lines were transduced with Lenti-Cas9-2A-Blast (Addgene, Cat# 73310) or Lenti-dCas9-KRAB-Blast (Addgene, Cat# 89567). .. MCF7 cells stably expressing Cas9 or KRAB-dCas9 were established by transduction of the virus at MOI < 0.7 , selected in 10 μg/ml blasticidin (Gibco, Cat# A1113903) and maintained in 5 μg/ml blasticidin.

    Article Title: CRISPR screens identify PRMT7 as a therapeutic target to enhance T cell-mediated killing in breast cancer.
    Article Snippet: .. Pooled CRISPR screens for resistance to T cell cytotoxicity For CRISPR screens and validations, breast cell lines were transduced with Lenti-Cas9-2A-Blast (Addgene, Cat# 73310) or Lenti-dCas9-KRAB-Blast (Addgene, Cat# 89567). .. MCF7 cells stably expressing Cas9 or KRAB-dCas9 were established by transduction of the virus at MOI<0.7 51, selected in 10 μg/ml blasticidin (Gibco, Cat# A1113903) and maintained in 5 μg/ml blasticidin.

    Article Title: HIV-1 transcription dominates over host gene activity at the HIV-1 integration site
    Article Snippet: .. For CRISPR-ready HIV-1-infected Jurkat T cell clones, HIV-1-infected Jurkat T cell clones (1G2, 8B10, 1D7, 1A8, 2F5, 5F9, and 1D9) were transduced with pseudotyped lentiviral vectors Lenti-MPHv2 (containing EF1α-driven MS2-p65-HSF1 activator and hygromycin selection marker, Addgene #89308) ( ) for CRISPRa and Lenti-dCas9-KRAB-blast (containing blasticidin selection marker, Addgene #89567) ( ) for CRISPRi, respectively. ..

    Plasmid Preparation:

    Article Title: BRRIAR lncRNA alters breast cancer risk by modulating interferon signaling in cis and in trans
    Article Snippet: Absorbance was measured at 450 nm using a Tecan i-control plate reader and concentrations calculated by interpolation using standard curves. .. Lenti-dCas9-KRAB-blast (a gift from Gary Hon [Addgene; #89567]), Lenti-Cas9‐2 A‐Blast plasmid (a gift from Jason Moffat [Addgene; #73310]), pgRNA-humanized (a gift from Stanley Qi [Addgene; #44248])or Human Brunello CRISPR knockout pooled library (a gift from David Root and John Doench [Addgene; #73178]) were co-transfected with two package plasmids, pMD2.G (a gift from Didier Trono [Addgene; #12259]) and psPAX2 (a gift from Didier Trono [Addgene; #12260]) into HEK293 cells using FuGENE6 (Promega). ..

    Article Title: Massively parallel functional profiling prioritizes breast cancer risk variants and implicates CCDC88C in ER-positive breast cancer
    Article Snippet: .. The data used for the analyses described in this manuscript were obtained from the GTEx API on April 20, 2026. pLS-SceI was gifted by Nadav Ahituv (Addgene plasmid #137725; http://n2t.net/addgene:137725 ; RRID:Addgene_137725), psPAX2 was gifted by Didier Trono (Addgene plasmid #12260; http://n2t.net/addgene:12260 ; RRID:Addgene_12260), pMD2.G was gifted by Didier Trono (Addgene plasmid #12259; http://n2t.net/addgene:12259 ; RRID:Addgene_12259), pKLV2-U6gRNA5(BbsI)-PGKpuro2AZsG-W was a gift from Kosuke Yusa (Addgene plasmid #67975; http://n2t.net/addgene:67975 ; RRID:Addgene_67975), and Lenti-dCas9-KRAB-blast was gifted by Gary Hon (Addgene plasmid #89567; http://n2t.net/addgene:89567 ; RRID:Addgene_89567). ..

    Knock-Out:

    Article Title: BRRIAR lncRNA alters breast cancer risk by modulating interferon signaling in cis and in trans
    Article Snippet: Absorbance was measured at 450 nm using a Tecan i-control plate reader and concentrations calculated by interpolation using standard curves. .. Lenti-dCas9-KRAB-blast (a gift from Gary Hon [Addgene; #89567]), Lenti-Cas9‐2 A‐Blast plasmid (a gift from Jason Moffat [Addgene; #73310]), pgRNA-humanized (a gift from Stanley Qi [Addgene; #44248])or Human Brunello CRISPR knockout pooled library (a gift from David Root and John Doench [Addgene; #73178]) were co-transfected with two package plasmids, pMD2.G (a gift from Didier Trono [Addgene; #12259]) and psPAX2 (a gift from Didier Trono [Addgene; #12260]) into HEK293 cells using FuGENE6 (Promega). ..

    Generated:

    Article Title: Multidimensional mapping of stimulation-responsive regulatory elements and candidate causal variants in T cell activation
    Article Snippet: .. Single-cell–derived stable Jurkat-dCas9-VP64 and Jurkat-dCas9-KRAB cell lines were generated using the Lenti-dCas9-VP64-blast (Addgene, 61425) and Lenti-dCas9-KRAB-blast (Addgene, 89567) vectors. .. Single-cell–derived stable Jurkat-dCas9-VP64 and Jurkat-dCas9-KRAB cell lines were generated using the Lenti-dCas9-VP64-blast (Addgene, 61425) and Lenti-dCas9-KRAB-blast (Addgene, 89567) vectors.

    Clone Assay:

    Article Title: HIV-1 transcription dominates over host gene activity at the HIV-1 integration site
    Article Snippet: .. For CRISPR-ready HIV-1-infected Jurkat T cell clones, HIV-1-infected Jurkat T cell clones (1G2, 8B10, 1D7, 1A8, 2F5, 5F9, and 1D9) were transduced with pseudotyped lentiviral vectors Lenti-MPHv2 (containing EF1α-driven MS2-p65-HSF1 activator and hygromycin selection marker, Addgene #89308) ( ) for CRISPRa and Lenti-dCas9-KRAB-blast (containing blasticidin selection marker, Addgene #89567) ( ) for CRISPRi, respectively. ..

    Selection:

    Article Title: HIV-1 transcription dominates over host gene activity at the HIV-1 integration site
    Article Snippet: .. For CRISPR-ready HIV-1-infected Jurkat T cell clones, HIV-1-infected Jurkat T cell clones (1G2, 8B10, 1D7, 1A8, 2F5, 5F9, and 1D9) were transduced with pseudotyped lentiviral vectors Lenti-MPHv2 (containing EF1α-driven MS2-p65-HSF1 activator and hygromycin selection marker, Addgene #89308) ( ) for CRISPRa and Lenti-dCas9-KRAB-blast (containing blasticidin selection marker, Addgene #89567) ( ) for CRISPRi, respectively. ..

    Marker:

    Article Title: HIV-1 transcription dominates over host gene activity at the HIV-1 integration site
    Article Snippet: .. For CRISPR-ready HIV-1-infected Jurkat T cell clones, HIV-1-infected Jurkat T cell clones (1G2, 8B10, 1D7, 1A8, 2F5, 5F9, and 1D9) were transduced with pseudotyped lentiviral vectors Lenti-MPHv2 (containing EF1α-driven MS2-p65-HSF1 activator and hygromycin selection marker, Addgene #89308) ( ) for CRISPRa and Lenti-dCas9-KRAB-blast (containing blasticidin selection marker, Addgene #89567) ( ) for CRISPRi, respectively. ..

    Expressing:

    Article Title: Generation of a comprehensive epigenomic atlas in clear cell renal cell carcinoma informs kidney cancer progression and heritability.
    Article Snippet: : CPCP-K2A) were delivered by BTX (Harvard Apparatus) electroporation. .. To generate stable 786-O-dCas9-KRAB expressing cell line the Lenti-dCas9-KRAB-blast (Addgene #89567) was used. .. Virus transduced cells were maintained for 2 weeks under blasticidin (10 μg/mL, Invivogen) selection.



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    A , Schematic of the analysis of DepMap genome-wide CRISPR-Cas9 screen data. Tumour cell lines from DepMap22Q2 release (PMID: 39468210) were classified as having either deleterious BRCA1 mutations (8 tumour cell lines), deleterious BRCA2 mutations (3 tumour cell lines) or no deleterious mutations in either BRCA1 or BRCA2 (1070 tumour cell lines). Using the class comparison tool ( https://depmap.org/portal/interactive/custom_analysis ) either BRCA1 or BRCA2 synthetic lethal genes were identified by comparing gene dependencies in either BRCA1 mutant or BRCA2 mutant tumour cell line cohorts to those in the cohort of tumour cell lines with no deleterious mutations in either BRCA1 or BRCA2. B , Volcano plots illustrating BRCA1 or C , BRCA2 synthetic lethal effects identified from DepMap analysis described in A . Negative effects (x-axis) indicate synthetic lethal effects, positive effects indicate synthetic viable effects. D - G . FANCM synthetic lethal effects identified in four genome-wide CRISPR-Cas9 screens. Violin plots of CRISPR-Cas9 screen data described in PMID: 40033056 are shown. Negative Z-scores represent synthetic lethal effects. CIP2A is shown as a positive control, being synthetic lethal in all four screens. FANCM is highlighted as a synthetic lethal in SUM149 and RPE TP53 −/− BRCA1 −/− screens (both BRCA1 -mutant) but not in <t>CAPAN1</t> nor DLD1 BRCA2 −/− screens (both BRCA2 -mutant). H , Schematic of the olaparib resistance screen in BRCA1- and BRCA2-degron cells. I , Rank-ordered dotplot representing genome-wide viability scores (RRA) under olaparib treatment in BRCA1- or J , BRCA2-depleted cells. In I and J , the significance threshold (FDR<0.001 and RRA<0.001) is shown as a dotted line, genes involved in DNA-end protection are shown as red circles, while Fancm/Rmi are shown as blue circles. K , Cell survival after 5 days of BRCA2-depletion and treatment with increasing concentrations of olaparib in Fancm -WT (grey) and Fancm -KO (blue) cells. The mean ±SD of 2 independent clones is represented. Statistical analysis: two-way ANOVA. L , Rank ordered Drug Effect Z-score data from a CRISPR-interference screen for PARP inhibitor resistance in human BRCA2 -mutant Capan1 cells. Capan1 cells were transduced with a genome-wide CRISPR interference sgRNA library and then exposed to 2000 nM olaparib for three weeks, a concentration that reduced the population size by 80 %, SF 20 (see Methods). Gene-centric Drug Effect Z scores from 18,788 genes are shown, with sgRNA targeting FANCM causing the most profound PARPi resistance-causing effect.
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    Image Search Results


    A , Schematic of the analysis of DepMap genome-wide CRISPR-Cas9 screen data. Tumour cell lines from DepMap22Q2 release (PMID: 39468210) were classified as having either deleterious BRCA1 mutations (8 tumour cell lines), deleterious BRCA2 mutations (3 tumour cell lines) or no deleterious mutations in either BRCA1 or BRCA2 (1070 tumour cell lines). Using the class comparison tool ( https://depmap.org/portal/interactive/custom_analysis ) either BRCA1 or BRCA2 synthetic lethal genes were identified by comparing gene dependencies in either BRCA1 mutant or BRCA2 mutant tumour cell line cohorts to those in the cohort of tumour cell lines with no deleterious mutations in either BRCA1 or BRCA2. B , Volcano plots illustrating BRCA1 or C , BRCA2 synthetic lethal effects identified from DepMap analysis described in A . Negative effects (x-axis) indicate synthetic lethal effects, positive effects indicate synthetic viable effects. D - G . FANCM synthetic lethal effects identified in four genome-wide CRISPR-Cas9 screens. Violin plots of CRISPR-Cas9 screen data described in PMID: 40033056 are shown. Negative Z-scores represent synthetic lethal effects. CIP2A is shown as a positive control, being synthetic lethal in all four screens. FANCM is highlighted as a synthetic lethal in SUM149 and RPE TP53 −/− BRCA1 −/− screens (both BRCA1 -mutant) but not in CAPAN1 nor DLD1 BRCA2 −/− screens (both BRCA2 -mutant). H , Schematic of the olaparib resistance screen in BRCA1- and BRCA2-degron cells. I , Rank-ordered dotplot representing genome-wide viability scores (RRA) under olaparib treatment in BRCA1- or J , BRCA2-depleted cells. In I and J , the significance threshold (FDR<0.001 and RRA<0.001) is shown as a dotted line, genes involved in DNA-end protection are shown as red circles, while Fancm/Rmi are shown as blue circles. K , Cell survival after 5 days of BRCA2-depletion and treatment with increasing concentrations of olaparib in Fancm -WT (grey) and Fancm -KO (blue) cells. The mean ±SD of 2 independent clones is represented. Statistical analysis: two-way ANOVA. L , Rank ordered Drug Effect Z-score data from a CRISPR-interference screen for PARP inhibitor resistance in human BRCA2 -mutant Capan1 cells. Capan1 cells were transduced with a genome-wide CRISPR interference sgRNA library and then exposed to 2000 nM olaparib for three weeks, a concentration that reduced the population size by 80 %, SF 20 (see Methods). Gene-centric Drug Effect Z scores from 18,788 genes are shown, with sgRNA targeting FANCM causing the most profound PARPi resistance-causing effect.

    Journal: bioRxiv

    Article Title: The FANCM-RMI1/2 complex promotes genomic instability and PARP inhibitor sensitivity in BRCA2-deficicient cells

    doi: 10.64898/2026.01.15.699681

    Figure Lengend Snippet: A , Schematic of the analysis of DepMap genome-wide CRISPR-Cas9 screen data. Tumour cell lines from DepMap22Q2 release (PMID: 39468210) were classified as having either deleterious BRCA1 mutations (8 tumour cell lines), deleterious BRCA2 mutations (3 tumour cell lines) or no deleterious mutations in either BRCA1 or BRCA2 (1070 tumour cell lines). Using the class comparison tool ( https://depmap.org/portal/interactive/custom_analysis ) either BRCA1 or BRCA2 synthetic lethal genes were identified by comparing gene dependencies in either BRCA1 mutant or BRCA2 mutant tumour cell line cohorts to those in the cohort of tumour cell lines with no deleterious mutations in either BRCA1 or BRCA2. B , Volcano plots illustrating BRCA1 or C , BRCA2 synthetic lethal effects identified from DepMap analysis described in A . Negative effects (x-axis) indicate synthetic lethal effects, positive effects indicate synthetic viable effects. D - G . FANCM synthetic lethal effects identified in four genome-wide CRISPR-Cas9 screens. Violin plots of CRISPR-Cas9 screen data described in PMID: 40033056 are shown. Negative Z-scores represent synthetic lethal effects. CIP2A is shown as a positive control, being synthetic lethal in all four screens. FANCM is highlighted as a synthetic lethal in SUM149 and RPE TP53 −/− BRCA1 −/− screens (both BRCA1 -mutant) but not in CAPAN1 nor DLD1 BRCA2 −/− screens (both BRCA2 -mutant). H , Schematic of the olaparib resistance screen in BRCA1- and BRCA2-degron cells. I , Rank-ordered dotplot representing genome-wide viability scores (RRA) under olaparib treatment in BRCA1- or J , BRCA2-depleted cells. In I and J , the significance threshold (FDR<0.001 and RRA<0.001) is shown as a dotted line, genes involved in DNA-end protection are shown as red circles, while Fancm/Rmi are shown as blue circles. K , Cell survival after 5 days of BRCA2-depletion and treatment with increasing concentrations of olaparib in Fancm -WT (grey) and Fancm -KO (blue) cells. The mean ±SD of 2 independent clones is represented. Statistical analysis: two-way ANOVA. L , Rank ordered Drug Effect Z-score data from a CRISPR-interference screen for PARP inhibitor resistance in human BRCA2 -mutant Capan1 cells. Capan1 cells were transduced with a genome-wide CRISPR interference sgRNA library and then exposed to 2000 nM olaparib for three weeks, a concentration that reduced the population size by 80 %, SF 20 (see Methods). Gene-centric Drug Effect Z scores from 18,788 genes are shown, with sgRNA targeting FANCM causing the most profound PARPi resistance-causing effect.

    Article Snippet: A positive selection, genome-wide CRISPR interference screen in Capan1 ( BRCA2 -mutant) cells was carried out in a dCas9-KRAB+ Capan1 derivative (generated using Lenti-dCas9-KRAB-blast, Addgene #89567), using the Weissman hCRISPRi-v2 sgRNA library (gift from Jonathan Weissman, Addgene #83969).

    Techniques: Genome Wide, CRISPR, Comparison, Mutagenesis, Positive Control, Clone Assay, Transduction, Concentration Assay