Generation and validation of CRISPR-engineered human natural killer cell lines for research and therapeutic applications

STAR Protoc. 2021 Oct 21;2(4):100874. doi: 10.1016/j.xpro.2021.100874. eCollection 2021 Dec 17.

Abstract

Cytotoxic natural killer cells kill tumors and infected cells. We carried out CRISPR-based gene editing and transcriptional regulation in hard-to-manipulate NK-92 cells. NK-92-based therapies were found to be safe and efficacious in preclinical studies of cancers. Here, we have pioneered the generation and validation of NK-92 cells constitutively expressing Cas9 or dCas9 for knockout (CRISPRko), transcriptional activation (CRISPRa), or transcriptional repression (CRISPRi) of genes. Our CRISPR-engineered NK-92 cell platforms can be modified for research and off-the-shelf therapeutic applications.

Keywords: CRISPR; Cancer; Cell Biology; Immunology; Molecular Biology.

Publication types

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

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • Cell Culture Techniques / methods*
  • Cell Line
  • Gene Editing / methods*
  • Humans
  • K562 Cells
  • Killer Cells, Natural / cytology*