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sabatini lander crispr pooled library  (Addgene inc)


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    Addgene inc sabatini lander crispr pooled library
    Sabatini Lander Crispr Pooled Library, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sabatini+lander+human+crispr+pooled+library/Human+Two+Plasmid+Activity-Optimized+CRISPR+Knockout+Library(Pooled+Library+%231000000095)/pm40801599-69-12-16
    Average 93 stars, based on 10 article reviews
    sabatini lander crispr pooled library - by Bioz Stars, 2026-09
    93/100 stars

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    CRISPR:

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.
    Article Snippet: The chromatin fraction was solubilized in Laemmli buffer plus β-mercaptoethanol and 1:100 Benzonase (Merck, #E1014). .. Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ). .. Oligonucleotides corresponding to top and bottom strands of the sgRNAs (Merck) were phosphorylated with T4 PNK (NEB, #M0201), denatured by heating to 95 ◦ C, annealed by cooling to 4 ◦ C at a rate of 0.1 ◦ C/s and then cloned into the lentiviral sgRNA expression vector pKLV2-U6gRNA(BbsI)-PGKpuro2ABFP-WPRE (Addgene #67974, kindly deposited by Kosuke Yusa 80 ).

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.
    Article Snippet: Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin. .. PRC2-reporter KBM-7 cells 34 expressing Cas9 were mutagenized using the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) at a multiplicity of infection of ∼0.3. .. Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin.

    Knock-Out:

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.
    Article Snippet: The chromatin fraction was solubilized in Laemmli buffer plus β-mercaptoethanol and 1:100 Benzonase (Merck, #E1014). .. Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ). .. Oligonucleotides corresponding to top and bottom strands of the sgRNAs (Merck) were phosphorylated with T4 PNK (NEB, #M0201), denatured by heating to 95 ◦ C, annealed by cooling to 4 ◦ C at a rate of 0.1 ◦ C/s and then cloned into the lentiviral sgRNA expression vector pKLV2-U6gRNA(BbsI)-PGKpuro2ABFP-WPRE (Addgene #67974, kindly deposited by Kosuke Yusa 80 ).

    Expressing:

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.
    Article Snippet: Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin. .. PRC2-reporter KBM-7 cells 34 expressing Cas9 were mutagenized using the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) at a multiplicity of infection of ∼0.3. .. Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin.

    Infection:

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.
    Article Snippet: Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin. .. PRC2-reporter KBM-7 cells 34 expressing Cas9 were mutagenized using the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) at a multiplicity of infection of ∼0.3. .. Untransduced KBM-7 cells were eliminated using either 500 μg/ml hygromycin (ThermoFisher Scientific, #10687010), 6.7 μg/ml blasticidin (Stratech, #B4879) or 0.75 μg/ml puromycin (ThermoFisher Scientific, #A1113803); untransduced K562 cells were eliminated using either 250 μg/ml hygromycin, 10 μg/ml blasticidin or 1 μg/ml puromycin.



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    Figure 1. A genome-wide <t>CRISPR-Cas9</t> genetic screen identifies an essential requirement for CRAMP1 and histone H1.4 in PRC2-mediated reporter repression (A) Schematic representation of GFP reporter repression by the PRC2 complex. (B) The GFP reporter is derepressed upon CRISPR-Cas9-mediated gene disruption of any of the three core PRC2 subunits, as assayed by flow cytometry. (C) A genome-wide CRISPR-Cas9 screen to identify factors required for PRC2 function. Following Cas9 expression in KBM-7 cells harboring the PRC2-sensitive GFP reporter, genome-wide mutagenesis was carried out with the Sabatini/Lander single guide RNA (sgRNA) library, 36 and GFP + cells isolated through two sequential rounds of FACS. ‘‘Significance’’ on the y axis represents the negative log of the ‘‘pos|score’’ metric reported by Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout (MAGeCK). 37
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    david sabatini eric lander  (Addgene inc)
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    Figure 1. A genome-wide <t>CRISPR-Cas9</t> genetic screen identifies an essential requirement for CRAMP1 and histone H1.4 in PRC2-mediated reporter repression (A) Schematic representation of GFP reporter repression by the PRC2 complex. (B) The GFP reporter is derepressed upon CRISPR-Cas9-mediated gene disruption of any of the three core PRC2 subunits, as assayed by flow cytometry. (C) A genome-wide CRISPR-Cas9 screen to identify factors required for PRC2 function. Following Cas9 expression in KBM-7 cells harboring the PRC2-sensitive GFP reporter, genome-wide mutagenesis was carried out with the Sabatini/Lander single guide RNA (sgRNA) library, 36 and GFP + cells isolated through two sequential rounds of FACS. ‘‘Significance’’ on the y axis represents the negative log of the ‘‘pos|score’’ metric reported by Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout (MAGeCK). 37
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    Image Search Results


    Figure 1. A genome-wide CRISPR-Cas9 genetic screen identifies an essential requirement for CRAMP1 and histone H1.4 in PRC2-mediated reporter repression (A) Schematic representation of GFP reporter repression by the PRC2 complex. (B) The GFP reporter is derepressed upon CRISPR-Cas9-mediated gene disruption of any of the three core PRC2 subunits, as assayed by flow cytometry. (C) A genome-wide CRISPR-Cas9 screen to identify factors required for PRC2 function. Following Cas9 expression in KBM-7 cells harboring the PRC2-sensitive GFP reporter, genome-wide mutagenesis was carried out with the Sabatini/Lander single guide RNA (sgRNA) library, 36 and GFP + cells isolated through two sequential rounds of FACS. ‘‘Significance’’ on the y axis represents the negative log of the ‘‘pos|score’’ metric reported by Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout (MAGeCK). 37

    Journal: Molecular cell

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.

    doi: 10.1016/j.molcel.2025.05.031

    Figure Lengend Snippet: Figure 1. A genome-wide CRISPR-Cas9 genetic screen identifies an essential requirement for CRAMP1 and histone H1.4 in PRC2-mediated reporter repression (A) Schematic representation of GFP reporter repression by the PRC2 complex. (B) The GFP reporter is derepressed upon CRISPR-Cas9-mediated gene disruption of any of the three core PRC2 subunits, as assayed by flow cytometry. (C) A genome-wide CRISPR-Cas9 screen to identify factors required for PRC2 function. Following Cas9 expression in KBM-7 cells harboring the PRC2-sensitive GFP reporter, genome-wide mutagenesis was carried out with the Sabatini/Lander single guide RNA (sgRNA) library, 36 and GFP + cells isolated through two sequential rounds of FACS. ‘‘Significance’’ on the y axis represents the negative log of the ‘‘pos|score’’ metric reported by Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout (MAGeCK). 37

    Article Snippet: Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ).

    Techniques: Genome Wide, CRISPR, Disruption, Flow Cytometry, Expressing, Mutagenesis, Isolation, Knock-Out

    Figure 5. Linker histones are not enriched at regions marked by H3K9me3 (A–D) Lack of linker histone enrichment at H3K9me3-marked genomic regions. (A) Tornado plots depicting linker histone CUT&Tag signal across H3K9me3 peaks from the ENCODE project; average signal intensity is shown in (B). (C) Heatmap depicting the lack of correlation between linker histone occupancy and H3K9me3. Cells are annotated with pairwise Spearman correlation coefficients. An example locus is shown in (D). (E) CUT&Tag faithfully profiles H3K9me3. Example loci comparing CUT&Tag versus H3K9me3 ChIP-seq data (ENCODE) are shown. (F and G) Linker histone insufficiency does not impair H3K9me3-dependent LINE-1 silencing by the HUSH complex. (F) Schematic representation of the dual- color reporter cell line designed to monitor both H3K9me3-dependent repression by the HUSH complex and linker histone-mediated PRC2-reporter repression. (G) HUSH-mediated LINE-1 silencing is unaffected upon CRAMP1 depletion. The indicated CRISPR sgRNAs were expressed in the dual-color reporter cell line, and GFP and iRFP fluorescence assayed by flow cytometry. See also Figure S5 and Table S2.

    Journal: Molecular cell

    Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.

    doi: 10.1016/j.molcel.2025.05.031

    Figure Lengend Snippet: Figure 5. Linker histones are not enriched at regions marked by H3K9me3 (A–D) Lack of linker histone enrichment at H3K9me3-marked genomic regions. (A) Tornado plots depicting linker histone CUT&Tag signal across H3K9me3 peaks from the ENCODE project; average signal intensity is shown in (B). (C) Heatmap depicting the lack of correlation between linker histone occupancy and H3K9me3. Cells are annotated with pairwise Spearman correlation coefficients. An example locus is shown in (D). (E) CUT&Tag faithfully profiles H3K9me3. Example loci comparing CUT&Tag versus H3K9me3 ChIP-seq data (ENCODE) are shown. (F and G) Linker histone insufficiency does not impair H3K9me3-dependent LINE-1 silencing by the HUSH complex. (F) Schematic representation of the dual- color reporter cell line designed to monitor both H3K9me3-dependent repression by the HUSH complex and linker histone-mediated PRC2-reporter repression. (G) HUSH-mediated LINE-1 silencing is unaffected upon CRAMP1 depletion. The indicated CRISPR sgRNAs were expressed in the dual-color reporter cell line, and GFP and iRFP fluorescence assayed by flow cytometry. See also Figure S5 and Table S2.

    Article Snippet: Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ).

    Techniques: ChIP-sequencing, CRISPR, Fluorescence, Flow Cytometry