Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells

View ORCID ProfilePinar Aksoy, View ORCID ProfileBülent Arman Aksoy, View ORCID ProfileEric Czech, View ORCID ProfileJeff Hammerbacher
doi: https://doi.org/10.1101/466243
Pinar Aksoy
*Microbiology and Immunology Department at the Medical University of South Carolina
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Pinar Aksoy
Bülent Arman Aksoy
*Microbiology and Immunology Department at the Medical University of South Carolina
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Bülent Arman Aksoy
Eric Czech
*Microbiology and Immunology Department at the Medical University of South Carolina
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Eric Czech
Jeff Hammerbacher
*Microbiology and Immunology Department at the Medical University of South Carolina
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Jeff Hammerbacher
  • For correspondence: hammer@hammerlab.org
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

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.

Footnotes

  • We added a new section showing how to express functional TCRs on mouse and human T cells by mRNA electroporation.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Back to top
PreviousNext
Posted October 20, 2019.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells
Pinar Aksoy, Bülent Arman Aksoy, Eric Czech, Jeff Hammerbacher
bioRxiv 466243; doi: https://doi.org/10.1101/466243
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells
Pinar Aksoy, Bülent Arman Aksoy, Eric Czech, Jeff Hammerbacher
bioRxiv 466243; doi: https://doi.org/10.1101/466243

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Immunology
Subject Areas
All Articles
  • Animal Behavior and Cognition (3585)
  • Biochemistry (7539)
  • Bioengineering (5494)
  • Bioinformatics (20724)
  • Biophysics (10292)
  • Cancer Biology (7946)
  • Cell Biology (11609)
  • Clinical Trials (138)
  • Developmental Biology (6584)
  • Ecology (10161)
  • Epidemiology (2065)
  • Evolutionary Biology (13573)
  • Genetics (9511)
  • Genomics (12811)
  • Immunology (7900)
  • Microbiology (19490)
  • Molecular Biology (7632)
  • Neuroscience (41969)
  • Paleontology (307)
  • Pathology (1254)
  • Pharmacology and Toxicology (2189)
  • Physiology (3258)
  • Plant Biology (7017)
  • Scientific Communication and Education (1293)
  • Synthetic Biology (1946)
  • Systems Biology (5417)
  • Zoology (1112)