Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells.

ABSTRACT: CRISPR (clustered, regularly interspaced, short palindromic repeats)/Cas9 (CRISPR-associated protein 9) has become the tool of choice for generating gene knockouts across a variety of species. The ability for efficient gene editing in primary T cells not only represents a valuable research tool to study gene function but also holds great promise for T cell-based immunotherapies, such as next-generation chimeric antigen receptor (CAR) T cells. Previous attempts to apply CRIPSR/Cas9 for gene editing in primary T cells have resulted in highly variable knockout efficiency and required T cell receptor (TCR) stimulation, thus largely precluding the study of genes involved in T cell activation or differentiation. Here, we describe an optimized approach for Cas9/RNP transfection of primary mouse a

SUBMITTER: Seki A 

PROVIDER: S-EPMC5839763 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature