Methods and Protocols (Nov 2020)

CRISPR-Based Editing Techniques for Genetic Manipulation of Primary T Cells

  • Mateusz Kotowski,
  • Sumana Sharma

DOI
https://doi.org/10.3390/mps3040079
Journal volume & issue
Vol. 3, no. 4
p. 79

Abstract

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While clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing techniques have been widely adapted for use in immortalised immune cells, efficient manipulation of primary T cells has proved to be more challenging. Nonetheless, the rapid expansion of the CRISPR toolbox accompanied by the development of techniques for delivery of CRISPR components into primary T cells now affords the possibility to genetically manipulate primary T cells both with precision and at scale. Here, we review the key features of the techniques for primary T cell editing and discuss how the new generation of CRISPR-based tools may advance genetic engineering of these immune cells. This improved ability to genetically manipulate primary T cells will further enhance our fundamental understanding of cellular signalling and transcriptional networks in T cells and more importantly has the potential to revolutionise T cell-based therapies.

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