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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.
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Depositing Labs
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Publication
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 | |
† 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
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SpeciesHuman
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Lentiviral Generation3rd
| 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 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.
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Volume∼20 µL
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Concentration10 ng/µL
Resource Information
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Protocols
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Depositor Data
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Terms and Licenses
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.
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.
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For your Materials & Methods section:
DNA Repair Library was a gift from Brittany Adamson (Addgene # ) -
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
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