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

用于扩增和 CRISPR-Cas9 基因组编辑小鼠 MAIT 细胞以进行后续体内研究的方案

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作者:Anastasia du Halgouet ,Aurélie Darbois ,Aurélia Alphonse ,Thomas Yvorra ,Ludovic Colombeau ,Raphaël Rodriguez ,Olivier Lantz ,Marion Salou

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.

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