Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells
Aksoy, P.; Aksoy, B. A.; Czech, E.; Hammerbacher, J.
Show abstract
Electroporation is the most feasible non-viral material delivery system for manipulating human T cells given its time- and cost-effectiveness. However, efficient delivery requires electroporation settings to be optimized for different devices, cellular states, and materials to be delivered. Here, we used electroporation to either induce exogenous gene expression in human primary T cells by plasmids or in vitro transcribed (IVT) mRNA and also target endogenous genes by Cas9 ribonucleoproteins (RNPs). We characterized the electroporation conditions both for activated and unstimulated human T cells. Although naive cells are non-dividing and therefore their genetic manipulation is harder compared to activated T cells, we developed the technical ability to manipulate both naive and memory cells within the unstimulated T cell population by IVT mRNA and Cas9 RNP electroporation. Here, we outline the best practices for achieving highly-efficient genetic manipulation in primary T cells without causing significant cytotoxicity to the cells. Because there is increasing evidence for \"less-differentiated\" T cells to have better anti-tumor activity for immunotherapy, manipulating naive T cells with high efficiency is also of high importance to clinical applications and to study the biology of these cells.
Matching journals
The top 11 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells 94%
- An optimized workflow for single-cell transcriptomics and repertoire profiling of purified lymphocytes from clinical samples. 92%
- Generating an organ-deficient animal model using a multi-targeted CRISPR-Cas9 system 92%
Similar papers in this journal
Similar papers in this journal
- Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system 91%
- Transient light-activated gene expression in Chinese hamster ovary cells 91%
- A simplified two-plasmid system for orthogonal control of gene expression using light-activated CRISPR effector for mammalian cells 90%
Similar papers in this journal
- CRISPR-Cas9 HDR Optimization: RAD52, Denatured and 5'-Modified DNA Templates in Knock-In Mice Generation 90%
- A long intergenic non-coding RNA regulates nuclear localisation of DNA methyl transferase-1 90%
- Identification of genome safe harbor loci for human gene therapy based on evolutionary biology and comparative genomics 90%
Similar papers in this journal
- Naive and in vitro-activated primary mouse CD8+ T cells retain in vivo immune responsiveness after electroporation-based CRISPR/Cas9 genetic engineering 93%
- Live-cell microscopy reveals that human T cells primarily respond chemokinetically within a CCL19 gradient that induces chemotaxis in dendritic cells 92%
- TNFa and IL-6 promote ex-vivo proliferation of lineage-committed human regulatory T cells 91%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.