High-efficiency nonviral CRISPR/Cas9-mediated gene editing of human T cells using plasmid donor DNA.
J Exp Med
; 219(5)2022 05 02.
Article
em En
| MEDLINE
| ID: mdl-35452075
ABSTRACT
Genome engineering of T lymphocytes, the main effectors of antitumor adaptive immune responses, has the potential to uncover unique insights into their functions and enable the development of next-generation adoptive T cell therapies. Viral gene delivery into T cells, which is currently used to generate CAR T cells, has limitations in regard to targeting precision, cargo flexibility, and reagent production. Nonviral methods for effective CRISPR/Cas9-mediated gene knock-out in primary human T cells have been developed, but complementary techniques for nonviral gene knock-in can be cumbersome and inefficient. Here, we report a convenient and scalable nonviral method that allows precise gene edits and transgene integration in primary human T cells, using plasmid donor DNA template and Cas9-RNP. This method is highly efficient for single and multiplex gene manipulation, without compromising T cell function, and is thus valuable for use in basic and translational research.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sistemas CRISPR-Cas
/
Edição de Genes
Limite:
Humans
Idioma:
En
Revista:
J Exp Med
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Canadá