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Inzolia Human CRISPR/Cas12a Multiplex Knockout Pooled Library
(Pooled Library #209551, #209552)

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  • Purpose

    This human genome-wide, CRISPR knockout enAsCas12a gRNA library targets single genes and paralog pairs, triples, and quads with arrays of four gRNAs. 19,687 single genes are targeted with two constructs, each of which has four guides, with the second construct encoding the same gRNAs in different order in the array. 4,435 paralog pairs are targeted by a single construct with two gRNAs per gene. 376 paralog triples are targeted by a single construct with one gRNA per gene and are padded with a fourth non-targeting guide to extend the 3-mer to a 4-mer array. Finally, 100 paralog quads are targeted by a single construct with one gRNA per gene. The library also contains 20 arrays targeting EGFP, 500 arrays targeting intergenic sites, and 50 arrays targeting non-targeting sites for a total of 49,766 arrays in the library.

  • Vector Backbone

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 209551 Inzolia library in pRDA_052† 1 $540 Add to Cart
Pooled Library 209552 Inzolia library in pRDA_550 1 $540 Add to Cart
Available to Academic and Nonprofits Only

  • † A Cas12 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pRDA_174 (Addgene #136476) or any other plasmid(s) or cell lines expressing enAsCas12a.

Library Details

  • Species
    Human
  • Genes targeted

    • 19,687 single genes
    • 4,435 paralog pairs
    • 376 paralog triples
    • 100 paralog quads
  • gRNAs

    • for single genes: 4 guides/gene/construct x 2 constructs (second construct has the same 4 guides in a different order in the array)
    • for paralog pairs: 2 guides/gene x 2 genes/construct
    • for paralog triples: 1 guide/gene x 3 genes/construct (non-targeting sequences added to extend 3mer paralog constructs into 4mer guide arrays)
    • for paralog quads: 1 guide/gene x 4 genes/construct
  • Controls

    • 20 arrays targeting EGFP
    • 500 arrays targeting intergenic site
    • 50 arrays targeting non-targeting sites
  • Total number of arrays
    49,766
  • Lentiviral Generation
    3rd

Library Shipping

This library is delivered as suspended DNA 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

Schematic showing two example arrays of 4 Cas12a gRNAs for each of the 4 categories: "19,687 single genes" (4 crRNAs targeting a single gene), "4,435 paralog pairs" (a pair of crRNAs targeting 2 genes), "376 paralog triples" (a triplet of crRNAs targeting 3 genes), and "100 paralog quads" (a quadruplet of crRNAs targeting 4 genes). Interspaced between the crRNAs are “DR4”, “DR3”, and “DR1” with "polyT" at the 3' end.

Figure 1: In4mer platform for whole-genome screening. Inzolia human whole-genome library targets single genes and paralog pairs, triples, and quads with arrays of four Cas12a gRNAs. Each gene or gene family is targeted by two arrays. Commercially-synthesized oligo pools were cloned into the one component pRDA_052 or pRDA_550 lentiviral vector. Image created in Biorender.

Please visit https://doi.org/10.1101/2023.01.03.522655 (Link opens in a new window) for bioRxiv preprint.

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:

    Inzolia CRISPR/Cas12a Multiplex Knockout Pooled Library in pRDA_052 was a gift from John Doench (Addgene #209551)
    Inzolia CRISPR/Cas12a Multiplex Knockout Pooled Library in pRDA_550 was a gift from John Doench (Addgene #209552)

  • For your References section:

    Efficient gene knockout and genetic interaction screening using the in4mer CRISPR/Cas12a multiplex knockout platform. Esmaeili Anvar N, Lin C, Ma X, Wilson LL, Steger R, Sangree AK, Colic M, Wang SH, Doench JG, Hart T. Nat Commun. 2024 Apr 27;15(1):3577. doi: 10.1038/s41467-024-47795-3. PubMed 38678031 (Link opens in a new window)
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