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

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...

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Detalles Bibliográficos
Autores principales: du Halgouet, Anastasia, Darbois, Aurélie, Alphonse, Aurélia, Yvorra, Thomas, Colombeau, Ludovic, Rodriguez, Raphaël, Lantz, Olivier, Salou, Marion
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10362169/
https://www.ncbi.nlm.nih.gov/pubmed/37432855
http://dx.doi.org/10.1016/j.xpro.2023.102419
Descripción
Sumario: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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