Generation of knock-in primary human T cells using Cas9 ribonucleoproteins
Abstract
T-cell genome engineering holds great promise for cancer immunotherapies and cell-based therapies for HIV, primary immune deficiencies, and autoimmune diseases, but genetic manipulation of human T cells has been inefficient. We achieved efficient genome editing by delivering Cas9 protein pre-assembled with guide RNAs. These active Cas9 ribonucleoproteins (RNPs) enabled successful Cas9-mediated homology-directed repair in primary human T cells. Cas9 RNPs provide a programmable tool to replace specific nucleotide sequences in the genome of mature immune cells—a longstanding goal in the field. These studies establish Cas9 RNP technology for diverse experimental and therapeutic genome engineering applications in primary human T cells.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- August 2015
- DOI:
- 10.1073/pnas.1512503112
- Bibcode:
- 2015PNAS..11210437S