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Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies.

ABSTRACT: Generating knockout mice for target molecules in specific T cell populations, without subset-specific promoters, is time-consuming and costly. Here, we describe steps for enriching mucosal-associated invariant T cells from the thymus, expanding them in vitro and performing a CRISPR-Cas9 knockout. We then detail procedure for injecting the knockout cells into wounded Cd3ε^-/- mice and characterizing them in the skin. For complete details on the use and execution of this protocol, please refer to du Halgouet et al. (2023).¹.

SUBMITTER: du Halgouet A

PROVIDER: S-EPMC10362169 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies.

du Halgouet Anastasia A Darbois Aurélie A Alphonse Aurélia A Yvorra Thomas T Colombeau Ludovic L Rodriguez Raphaël R Lantz Olivier O Salou Marion M

STAR protocols 20230709 3

Generating knockout mice for target molecules in specific T cell populations, without subset-specific promoters, is time-consuming and costly. Here, we describe steps for enriching mucosal-associated invariant T cells from the thymus, expanding them in vitro and performing a CRISPR-Cas9 knockout. We then detail procedure for injecting the knockout cells into wounded Cd3ε<sup>-/-</sup> mice and characterizing them in the skin. For complete details on the use and execution of this protocol, please ...[more]

PMID: 37432855

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