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DNA Repair CRISPRi Libraries
(Pooled Libraries #177663, #177664, #177670)

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  • Purpose

    These CRISPR-interference (CRISPRi) libraries contain sgRNAs targeting genes encoding factors involved in DNA repair and associated processes. These libraries can be used in Repair-seq experiments for mapping DNA repair pathways and identifying strategies to optimize genome editing. The backbone of #177663 and #177664 includes a Cas9 or Cas12a targeted site, while #177670 includes a prime editing targeted site.

    To perform Repair-seq experiments, transduce the CRISPRi library into cells expressing dCas-KRAB to deplete the target genes, then electroporate with ribonucleoprotein, plasmid, or mRNA of Cas9, Cas12, or prime editor with gRNA targeting the editing site on the library backbone.

  • Vector Backbone

    #177663 and #177664: pAX198 (Plasmid #173042) - does not express Cas9

    #177670: pPC1000 (Plasmid #174829) - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 177663 DNA Repair Library with 1,573 gRNAs† 1 $380 Add to Cart
Pooled Library 177664 DNA Repair Library with 336 gRNAs† 1 $380 Add to Cart
Pooled Library 177670 DNA Repair Library with 1,573 gRNAs (PE target)† 1 $380 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 pHR-SFFV-dCas9-BFP-KRAB (Addgene #46911) or other plasmid(s) or cell lines expressing dCas9-KRAB.

Library Details

  • Species
    Human
  • Lentiviral Generation
    3rd
Pooled Library Genes Targeted Targeting gRNAs Control gRNAs
#177663 476 1,573 60
#177664 118* 366 30
#177670 476 1,573 60

*The 118 genes targeted by #177664 are mostly, but not exclusively, a subset of the 476 genes targeted by the larger libraries.

Library Shipping

Each 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
    ∼20 µL
  • Concentration
    10 ng/µL

Resource Information

Depositor Comments

More information, including interactive exploration of raw data, is available in the Repair-seq data browser (Link opens in a new window). Analysis scripts are available at GitHub (Link opens in a new window).

It is also recommended to PCR and sequence the initial library upon receipt. Note that we introduced the skewness in the library with 336 sgRNAs intentionally, to compensate for the loss of some toxic guides.

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:

    DNA Repair CRISPRi Library with 1,573 gRNAs was a gift from Brittany Adamson (Addgene #177663)
    DNA Repair CRISPRi Library with 336 gRNAs was a gift from Brittany Adamson (Addgene #177664)
    DNA Repair CRISPRi Library with 1,573 gRNAs (PE target) was a gift from Brittany Adamson (Addgene #177670)

  • For your References section:

    For #177663 and #177664:

    Mapping the genetic landscape of DNA double-strand break repair. Hussmann JA, Ling J, Ravisankar P, Yan J, Cirincione A, Xu A, Simpson D, Yang D, Bothmer A, Cotta-Ramusino C, Weissman JS, Adamson B. Cell. 2021 Oct 28;184(22):5653-5669.e25. doi: 10.1016/j.cell.2021.10.002. PubMed 34672952 (Link opens in a new window)

    For #177670:

    Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. PubMed 34653350 (Link opens in a new window)
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