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crispr interference crispri screening approach  (Addgene inc)


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    Addgene inc crispr interference crispri screening approach
    Fig. 5. <t>CRISPRi</t> screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).
    Crispr Interference Crispri Screening Approach, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/crispr+interference+crispri+screening+approach/pm40184725-195-31-46?v=Addgene+inc
    Average 94 stars, based on 55 article reviews
    crispr interference crispri screening approach - by Bioz Stars, 2026-07
    94/100 stars

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    1) Product Images from "A quiescence-like/TGF-β1-specific CRISPRi screen reveals drug uptake transporters as secondary targets of kinase inhibitors in AML."

    Article Title: A quiescence-like/TGF-β1-specific CRISPRi screen reveals drug uptake transporters as secondary targets of kinase inhibitors in AML.

    Journal: Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy

    doi: 10.1016/j.drup.2025.101242

    Fig. 5. CRISPRi screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).
    Figure Legend Snippet: Fig. 5. CRISPRi screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).

    Techniques Used: Transduction, Amplification, Biomarker Discovery, Competitive Binding Assay, Flow Cytometry



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    crispr interference crispri screening approach  (Addgene inc)
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    Addgene inc crispr interference crispri screening approach
    Fig. 5. <t>CRISPRi</t> screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).
    Crispr Interference Crispri Screening Approach, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/crispr+interference+crispri+screening+approach/pm40184725-195-31-46?v=Addgene+inc
    Average 94 stars, based on 1 article reviews
    crispr interference crispri screening approach - by Bioz Stars, 2026-07
    94/100 stars
    		  Buy from Supplier

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    Fig. 5. CRISPRi screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).

    Journal: Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy

    Article Title: A quiescence-like/TGF-β1-specific CRISPRi screen reveals drug uptake transporters as secondary targets of kinase inhibitors in AML.

    doi: 10.1016/j.drup.2025.101242

    Figure Lengend Snippet: Fig. 5. CRISPRi screening reveals a potential link between vactosertib-induced resistance to Ara-C and alterations in drug uptake mechanisms in AML cells. (A) Schematic of the CRISPRi library screen workflow. THP1-dCas9-KRAB cells were transduced with CRISPRi library at an MOI of approximately 0.3 and selected with puromycin. Cells were treated with either Ara-C alone or in combination with vactosertib for 48 hours and were maintained for 21 days in culture. sgRNA sequences were amplified from extracted DNA and analyzed by NGS. MAGeCK was used for data processing. (B) and (C) Volcano plots indicating sgRNA distribution in vactosertib + Ara-C and Ara-C alone conditions, respectively. Data represent three independent biological replicates (n = 3). For every gene, the x-axis displays its enrichment (positive hits) or depletion (negative hits) after treatment, while the y-axis shows the score as calculated by MAGeCK. Relevant hits were annotated. (D) Validation of CRISPRi library screen results by cell growth competition assay. THP1-dCas9-KRAB cells were transduced with sgENT1 (two independent sgRNAs: sgENT1#1 and sgENT1#2), sgDCK, sgNT and sgMYBL2. The effect of each sgRNA on cell proliferation under different treatment conditions with vactosertib and Ara- C was determined by tracking the percentage of BFP-positive cells over time using flow cytometry. Results are shown as the percentage of BFP-positive cells at the indicated times relative to day 0 (n = 3).

    Article Snippet: A drug-focused CRISPR interference (CRISPRi) library screen reveals TGFBR1 as the primary mediator of the quiescence-like phenotype induced by TGF-β1 To explore potential therapeutic interventions targeting TGF-β1induced dormancy, we employed a CRISPR interference (CRISPRi) screening approach using a sgRNA library targeting kinases, phosphatases and drug targets (Addgene #83971) (Horlbeck et al., 2016).

    Techniques: Transduction, Amplification, Biomarker Discovery, Competitive Binding Assay, Flow Cytometry