Optimized protocol to create deletion in adherent cell lines using CRISPR/Cas9 system

STAR Protoc. 2021 Oct 22;2(4):100857. doi: 10.1016/j.xpro.2021.100857. eCollection 2021 Dec 17.


The CRISPR/Cas9 system is a powerful tool for genome editing and is adaptable for a wide range of applications. Here, we have put together a step-by-step protocol for generating knockout cell lines (coding or non-coding region) using CRISPR/Cas9 tool. The protocol below has been tested on adherent cell lines such as HeLa and MCF7. However, it may easily be adapted to other adherent cell lines with minor variations. For complete details on the use and execution of this protocol, please refer to Farooq et al. (2021).

Keywords: CRISPR; Gene Expression; Genomics; Molecular Biology; Single Cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • Gene Deletion*
  • Gene Editing / methods*
  • HeLa Cells
  • Humans
  • MCF-7 Cells