Base Editing Protocol: Disruption of PDCD1, TIGIT, and TRAC Loci in Primary T Cells Using Adenine and Cytosine Base Editors Delivered as IVT mRNA v1

2026 Preprint Open access

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Authors: Jethro Langley, J.E. Curry, Lou Baudrier, Pierre Billon
DOI: 10.17504/protocols.io.rm7vzwmx8vx1/v1
Cited by: 1

Abstract

Targeted gene knockout of PDCD1 (PD-1), TIGIT, and TRAC in activated primary human T cells via adenine base editing (ABE) or cytosine base editing (CBE) to disrupt splice donor/acceptor sites. Base editors are delivered as in vitro-transcribed (IVT) mRNA together with chemically modified synthetic gRNAs by Neon electroporation. This protocol generates mock-edited or multiplex base-edited CAR-T cells following lentiviral CAR transduction.

Fields of Study & Subjects

CRISPR and Genetic Engineering CAR-T cell therapy research Virus-based gene therapy research

Publication details

Publisher
Springer Science and Business Media LLC
Publication date
28 May 2026
Document type
Preprint