Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins.

ABSTRACT: CRISPR/Cas9 technology is accelerating genome engineering in many cell types, but so far, gene delivery and stable gene modification have been challenging in primary NK cells. For example, transgene delivery using lentiviral or retroviral transduction resulted in a limited yield of genetically-engineered NK cells due to substantial procedure-associated NK cell apoptosis. We describe here a DNA-free method for genome editing of human primary and expanded NK cells using Cas9 ribonucleoprotein complexes (Cas9/RNPs). This method allowed efficient knockout of the TGFBR2 and HPRT1 genes in NK cells. RT-PCR data showed a significant decrease in gene expression level, and a cytotoxicity assay of a representative cell product suggested that the RNP-modified NK cells became less sensitive to TGFβ. G

SUBMITTER: Naeimi Kararoudi M 

PROVIDER: S-EPMC6101749 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature