Highly efficient multiplex human T cell engineering without double-strand breaks using Cas9 base editors

Beau R. Webber, Cara lin Lonetree, Mitchell G. Kluesner, Matthew J. Johnson, Emily J. Pomeroy, Miechaleen D. Diers, Walker S. Lahr, Garrett M. Draper, Nicholas J. Slipek, Branden S. Smeester, Klaus N. Lovendahl, Amber N. McElroy, Wendy R. Gordon, Mark J. Osborn, Branden S. Moriarity

Research output: Contribution to journalArticle

Abstract

The fusion of genome engineering and adoptive cellular therapy holds immense promise for the treatment of genetic disease and cancer. Multiplex genome engineering using targeted nucleases can be used to increase the efficacy and broaden the application of such therapies but carries safety risks associated with unintended genomic alterations and genotoxicity. Here, we apply base editor technology for multiplex gene modification in primary human T cells in support of an allogeneic CAR-T platform and demonstrate that base editor can mediate highly efficient multiplex gene disruption with minimal double-strand break induction. Importantly, multiplex base edited T cells exhibit improved expansion and lack double strand break-induced translocations observed in T cells edited with Cas9 nuclease. Our findings highlight base editor as a powerful platform for genetic modification of therapeutically relevant primary cell types.

Original languageEnglish (US)
Article number5222
JournalNature communications
Volume10
Issue number1
DOIs
StatePublished - Dec 1 2019

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Cell engineering
Cell Engineering
T-cells
strands
nuclease
genome
Genes
engineering
T-Lymphocytes
genes
therapy
platforms
Genome
Inborn Genetic Diseases
safety
induction
fusion
cancer
Technology
Safety

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

Cite this

Highly efficient multiplex human T cell engineering without double-strand breaks using Cas9 base editors. / Webber, Beau R.; Lonetree, Cara lin; Kluesner, Mitchell G.; Johnson, Matthew J.; Pomeroy, Emily J.; Diers, Miechaleen D.; Lahr, Walker S.; Draper, Garrett M.; Slipek, Nicholas J.; Smeester, Branden S.; Lovendahl, Klaus N.; McElroy, Amber N.; Gordon, Wendy R.; Osborn, Mark J.; Moriarity, Branden S.

In: Nature communications, Vol. 10, No. 1, 5222, 01.12.2019.

Research output: Contribution to journalArticle

Webber, Beau R. ; Lonetree, Cara lin ; Kluesner, Mitchell G. ; Johnson, Matthew J. ; Pomeroy, Emily J. ; Diers, Miechaleen D. ; Lahr, Walker S. ; Draper, Garrett M. ; Slipek, Nicholas J. ; Smeester, Branden S. ; Lovendahl, Klaus N. ; McElroy, Amber N. ; Gordon, Wendy R. ; Osborn, Mark J. ; Moriarity, Branden S. / Highly efficient multiplex human T cell engineering without double-strand breaks using Cas9 base editors. In: Nature communications. 2019 ; Vol. 10, No. 1.
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