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

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

doi:10.1016/j.xpro.2023.102419

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

STAR Protoc. 2023 Sep 15;4(3):102419. doi: 10.1016/j.xpro.2023.102419. Epub 2023 Jul 9.

Authors

Anastasia du Halgouet¹, Aurélie Darbois², Aurélia Alphonse², Thomas Yvorra³, Ludovic Colombeau³, Raphaël Rodriguez³, Olivier Lantz⁴, Marion Salou⁵

Affiliations

¹ INSERM U932, PSL University, Institut Curie, 75005 Paris, France; National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
² INSERM U932, PSL University, Institut Curie, 75005 Paris, France.
³ Institut Curie, CNRS, INSERM, PSL Research University, Equipe Labellisée Ligue Contre le Cancer, Paris, France.
⁴ INSERM U932, PSL University, Institut Curie, 75005 Paris, France; Laboratoire d'Immunologie Clinique, Institut Curie, Paris 75005, France; Centre d'Investigation Clinique en Biothérapie Gustave-Roussy Institut Curie (CIC-BT1428) Institut Curie, Paris 75005, France. Electronic address: olivier.lantz@curie.fr.
⁵ INSERM U932, PSL University, Institut Curie, 75005 Paris, France. Electronic address: marion.salou@curie.fr.

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.

Keywords: CRISPR; Cell culture; Immunology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems / genetics
Genomics
Mice
Mice, Knockout
Mucosal-Associated Invariant T Cells*
Promoter Regions, Genetic