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Human CRISPR Knockout Pooled Libraries (Jacquere)
(Pooled Library #247026, #247027)

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  • Purpose

    This is a human genome-wide CRISPR knockout library for the use of Streptococcus pyogenes Cas9. Each gene is targeted by three constructs. The design of this library is agnostic to the cell line in use.

    The Jacquere library is enriched for effective guides through prioritization of guides with strong on-target activity as predicted with the Rule Set 3 Sequence + Target Score (DeWeirdt et al., 2022 (Link opens in a new window)), strategic exclusion of guides with excessive off-target activity, and avoidance of target sites with high variant frequencies in the human population according to the gnomAD (Link opens in a new window) database. Further, Jacquere is designed against current gene annotations sourced from GENCODE (v47), RefSeq (v2024_08), and CHESS (v3.1.3), thereby offering coverage of the comprehensive and contemporary human genome.

    Pooled library #247026 is a split-vector library, with Cas9 and gRNAs supplied on different plasmids. Pooled library #247027 is an all-in-one library, with Cas9 and gRNAs on the same plasmid.

  • Vector Backbone

    Pooled Library #247026 (split-vector): pMV_AA050 (Plasmid #216107) - does not express Cas9

    Pooled Library #247027 (all-in-one): pRDA_734 (Plasmid #216097) - expresses Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 247026 Human CRISPR KO Library in pMV_AA050† 1 $594 Add to Cart
Pooled Library 247027 Human CRISPR KO Library in pRDA_734 1 $594 Add to Cart
Available to Academic and Nonprofits Only

  • † A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pMV_AA088 (Addgene #216104) or any other plasmid(s) or cell lines expressing Cas9.

Library Details

  • Species
    Human
  • Genes targeted
    20,550
  • gRNAs per gene
    3
  • Total gRNAs
    60,550
  • Controls
    900 intergenic-targeting, 100 non-targeting
  • Lentiviral generation
    3rd

Library Shipping

Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

  • Volume
    ∼25 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

Deconvolution algorithms for analyzing NGS data: Extract guide sequences from sequencing reads by running PoolQ with the search prefix “CACCG”, and count reads by alignment to a reference file of all possible guides present in the library.

Table column headings denote the source catalog of the genes: Total, GENCODE v47, RefSeq 08-2024, CHESS 3.1.3. The top row denotes number targeted by more than three guides: 2177, 2177, 0, 0. The next row denotes number targeted by 3 guides: 18235, 17709, 508, 18. The next row denotes number targeted by 2 guides: 88, 68, 20, 0. The next row denotes number targeted by 1 guide: 50, 31, 19, 0. The last row denoted the number not targetable: 162, 126, 40, 0.
Figure 1: Genes targeted in Jacquere, stratified by source catalog.
How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    Human CRISPR Knockout Pooled Library (Jacquere) split-vector was a gift from John Doench and David Root (Addgene #247026 ; http://n2t.net/addgene:247026 ; RRID:Addgene_247026)
    Human CRISPR Knockout Pooled Library (Jacquere) all-in-one was a gift from John Doench and David Root (Addgene #247027 ; http://n2t.net/addgene:247027 ; RRID:Addgene_247027)

  • For your References section:

    Balancing off-target and on-target considerations for optimized Cas9 CRISPR knockout library design. Drepanos LM, Srikanth S, Kaplan EG, Shah ST, Escude Velasco B, Merzouk S, Doench JG. bioRxiv 2025.08.26.672375. https://doi.org/10.1101/2025.08.26.672375
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