Girish, Vishruth, Sheltzer, Jason M (July 2020) A CRISPR Competition Assay to Identify Cancer Genetic Dependencies. Bio-protocol, 10 (14). e3682. ISSN 2331-8325
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 diseases & disorders Investigative techniques and equipment Investigative techniques and equipment > CRISPR-Cas9 |
| CSHL Authors: | |
| Communities: | CSHL Cancer Center Program > Cancer Genetics and Genomics Program CSHL labs > Sheltzer lab CSHL Cancer Center Program CSHL Cancer Center Shared Resources > Flow Cytometry Service |
| SWORD Depositor: | CSHL Elements |
| Depositing User: | CSHL Elements |
| Date: | 20 July 2020 |
| Date Deposited: | 26 Apr 2021 19:43 |
| Last Modified: | 13 Feb 2024 21:19 |
| PMCID: | PMC7842800 |
| URI: | https://repository.cshl.edu/id/eprint/39931 |
Actions (login required)
 |
Administrator's edit/view item |
