A CRISPR Competition Assay to Identify Cancer Genetic Dependencies.

Girish, Vishruth, Sheltzer, Jason M (July 2020) A CRISPR Competition Assay to Identify Cancer Genetic Dependencies. Bio-protocol, 10 (14). e3682. ISSN 2331-8325

URL: https://www.ncbi.nlm.nih.gov/pubmed/33659353
DOI: 10.21769/BioProtoc.3682

Abstract

The CRISPR/Cas9 system is a powerful tool for genome editing, wherein the RNA-guided nuclease Cas9 can be directed to introduce double-stranded breaks (DSBs) at a targeted locus. In mammalian cells, these DSBs are typically repaired through error-prone processes, resulting in insertions or deletions (indels) at the targeted locus. Researchers can use these Cas9-mediated lesions to probe the consequences of loss-of-function perturbations in genes of interest. Here, we describe an optimized protocol to identify specific genes required for cancer cell fitness through a CRISPR-mediated cellular competition assay. Identifying these genetic dependencies is of utmost importance, as they provide potential targets for anti-cancer drug development. This protocol provides researchers with a robust and scalable approach to investigate gene dependencies in a variety of cell lines and cancer types and to validate the results of high-throughput or whole-genome screens.

Item Type: Paper
Subjects: diseases & disorders > cancer
Investigative techniques and equipment > CRISPR-Cas9
CSHL Authors:
Communities: CSHL labs > Sheltzer lab
CSHL Cancer Center Program > Cancer Genetics and Genomics Program
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 20 July 2020
Date Deposited: 26 Apr 2021 19:43
Last Modified: 13 Jul 2021 16:20
PMCID: PMC7842800
URI: https://repository.cshl.edu/id/eprint/39931

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