One-step CRISPR-Cas9-mediated knockout of native TCRαß genes in human T cells using RNA electroporation.
STAR Protoc
; 4(1): 102112, 2023 03 17.
Article
em En
| MEDLINE
| ID: mdl-36853667
ABSTRACT
To avoid mispairing between native and introduced T cell receptors (TCRs) and to prevent graft-versus-host disease in allogeneic T cell therapies, TCRα and TCRß chains of native TCRs are knocked out via CRISPR-Cas9. We demonstrate the isolation and activation of CD8+ T cells followed by electroporation of T cells with in vitro transcribed eSpCas9(1.1)-P2A-EGFP mRNA and single-guide RNAs targeting the TCRα and TCRß constant regions. We then describe a flow cytometric analysis to determine TCR knockout efficiency.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Receptores de Antígenos de Linfócitos T alfa-beta
/
Linfócitos T CD8-Positivos
Limite:
Humans
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article