Polymer-stabilized Cas9 nanoparticles and modified repair templates increase genome editing efficiency.

ABSTRACT: Versatile and precise genome modifications are needed to create a wider range of adoptive cellular therapies^1-5. Here we report two improvements that increase the efficiency of CRISPR-Cas9-based genome editing in clinically relevant primary cell types. Truncated Cas9 target sequences (tCTSs) added at the ends of the homology-directed repair (HDR) template interact with Cas9 ribonucleoproteins (RNPs) to shuttle the template to the nucleus, enhancing HDR efficiency approximately two- to fourfold. Furthermore, stabilizing Cas9 RNPs into nanoparticles with polyglutamic acid further improves editing efficiency by approximately twofold, reduces toxicity, and enables lyophilized storage without loss of activity. Combining the two improvements increases gene targeting efficiency even at

SUBMITTER: Nguyen DN 

PROVIDER: S-EPMC6954310 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature