STAR Protocols (Dec 2021)

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

  • Anil Kumar,
  • Sung June Lee,
  • Qiao Liu,
  • Anthony K.N. Chan,
  • Sheela Pangeni Pokharel,
  • Jianhua Yu,
  • Chun-Wei Chen,
  • Srividya Swaminathan

Journal volume & issue
Vol. 2, no. 4
p. 100874

Abstract

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Summary: 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.

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