Chemical-genetic CRISPR-Cas9 screens in human cells using a pathway-specific library

STAR Protoc. 2021 Jul 28;2(3):100685. doi: 10.1016/j.xpro.2021.100685. eCollection 2021 Sep 17.

Abstract

The development of CRISPR-Cas9 screening techniques coupled with chemical inhibition of specific biological processes enables high-throughput investigation into many areas of molecular biology. We present a protocol to conduct ubiquitin proteasome system-specific chemical-genetic CRISPR-Cas9 screens in the human HAP1 cell line. This protocol can be adapted for use in other cell lines, with other compounds and types of treatments, and with any other sgRNA library. For complete details on the use and execution of this protocol, please refer to Hundley et al. (2021).

Keywords: CRISPR; Genetics; High Throughput Screening.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Cell Line
  • Gene Editing / methods*
  • Gene Library
  • Genetic Testing / methods*
  • High-Throughput Screening Assays / methods*
  • Humans
  • RNA, Guide, CRISPR-Cas Systems / genetics

Substances

  • RNA, Guide, CRISPR-Cas Systems