PT  - JOURNAL ARTICLE
AU  - Pinar Aksoy
AU  - Bülent Arman Aksoy
AU  - Eric Czech
AU  - Jeff Hammerbacher
TI  - Electroporation characteristics of human primary T cells
AID  - 10.1101/466243
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 466243
4099  - http://biorxiv.org/content/early/2019/03/27/466243.short
4100  - http://biorxiv.org/content/early/2019/03/27/466243.full
AB  - Genetic manipulation of primary human T cells is a valuable technique for basic research in immunology to explore gene function and clinical applications that involve T cells. Among other techniques such as viral-based methods, electroporation is the most feasible material delivery system for manipulating human T cells. In this study, we used electroporation to either induce exogenous gene expression in human primary T cells by plasmids and in vitro transcribed (IVT) mRNA or target endogenous genes by Cas9 ribonucleoproteins (RNPs). We characterized the electroporation conditions both for activated and unstimulated T cells. Although, naive cells are non-dividing and their metabolism is slower compared to activated T cells; we were able to manipulate both naive and memory cells within the unstimulated T cell population by IVT mRNA- and Cas9 RNP-electroporation with more than 95% and 80% efficiency, respectively, and by plasmids with more than 50% efficiency. Here, we outline the best practices for achieving highly-efficient and non-viral genetic manipulation in primary T cells without causing significant cytotoxicity to the cells. Because there is increasing evidence for younger cells to have better anti-tumor activity for immunotherapy, manipulating naïve T cells with high efficiency is of high importance to clinical applications.