Optimized two-step electroporation process to achieve efficient nonviral-mediated gene insertion into primary T cells.

ABSTRACT: The development of gene editing technologies over the past years has allowed the precise and efficient insertion of transgenes into the genome of various cell types. Knock-in approaches using homology-directed repair and designer nucleases often rely on viral vectors, which can considerably impact the manufacturing cost and timeline of gene-edited therapeutic products. An attractive alternative would be to use naked DNA as a repair template. However, such a strategy faces challenges such as cytotoxicity from double-stranded DNA (dsDNA) to primary cells. Here, we sought to study the kinetics of transcription activator-like effector nuclease (TALEN)-mediated gene editing in primary T cells to improve nonviral gene knock-in. Harnessing this knowledge, we developed a rapid and efficient gene i

SUBMITTER: Yang M 

PROVIDER: S-EPMC8727936 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature