Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary T Cells.

ABSTRACT: Adoptive cell transfer is an important approach for basic research and emerges as an effective treatment for various diseases, including infections and blood cancers. Direct genetic manipulation of primary immune cells opens up unprecedented research opportunities and could be applied to enhance cellular therapeutic products. In this article, we report highly efficient genome engineering in primary murine T cells using a plasmid-based RNA-guided CRISPR system. We developed a straightforward approach to ablate genes in up to 90% of cells and to introduce precisely targeted single nucleotide polymorphisms in up to 25% of the transfected primary T cells. We used gene editing-mediated allele switching to quantify homology-directed repair, systematically optimize experimental parameters, and ma

SUBMITTER: Kornete M 

PROVIDER: S-EPMC5857648 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature