Enrichment of transgene integrations by transient CRISPR activation of a silent reporter gene.

ABSTRACT: CRISPR-Cas-mediated site-specific integration of transgenes by homology-directed repair (HDR) is challenging, especially in primary cells, where inferior editing efficiency may impede the development of gene- and cellular therapies. Various strategies for enrichment of cells with transgene integrations have been developed, but most strategies either generate unwanted genomic scars or rely on permanent integration and expression of a reporter gene used for selection. However, stable expression of a reporter gene may perturb cell homeostasis and function. Here we develop a broadly applicable and versatile enrichment strategy by harnessing the capability of CRISPR activation (CRISPRa) to transiently induce expression of a therapeutically relevant reporter gene used for immunomagnetic enrichme

SUBMITTER: Mikkelsen NS 

PROVIDER: S-EPMC10009645 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature