Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies
Summary: 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 kno...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723003866 |
| _version_ | 1797783591421214720 |
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| author | Anastasia du Halgouet Aurélie Darbois Aurélia Alphonse Thomas Yvorra Ludovic Colombeau Raphaël Rodriguez Olivier Lantz Marion Salou |
| author_facet | Anastasia du Halgouet Aurélie Darbois Aurélia Alphonse Thomas Yvorra Ludovic Colombeau Raphaël Rodriguez Olivier Lantz Marion Salou |
| author_sort | Anastasia du Halgouet |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-13T00:28:03Z |
| format | Article |
| id | doaj.art-62d48bb00cef4386ac2793eaa83838bc |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-13T00:28:03Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-62d48bb00cef4386ac2793eaa83838bc2023-07-11T04:06:41ZengElsevierSTAR Protocols2666-16672023-09-0143102419Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studiesAnastasia du Halgouet0Aurélie Darbois1Aurélia Alphonse2Thomas Yvorra3Ludovic Colombeau4Raphaël Rodriguez5Olivier Lantz6Marion Salou7INSERM U932, PSL University, Institut Curie, 75005 Paris, France; National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USAINSERM U932, PSL University, Institut Curie, 75005 Paris, FranceINSERM U932, PSL University, Institut Curie, 75005 Paris, FranceInstitut Curie, CNRS, INSERM, PSL Research University, Equipe Labellisée Ligue Contre le Cancer, Paris, FranceInstitut Curie, CNRS, INSERM, PSL Research University, Equipe Labellisée Ligue Contre le Cancer, Paris, FranceInstitut Curie, CNRS, INSERM, PSL Research University, Equipe Labellisée Ligue Contre le Cancer, Paris, FranceINSERM 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; Corresponding authorINSERM U932, PSL University, Institut Curie, 75005 Paris, France; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723003866Cell cultureImmunologyCRISPR |
| spellingShingle | Anastasia du Halgouet Aurélie Darbois Aurélia Alphonse Thomas Yvorra Ludovic Colombeau Raphaël Rodriguez Olivier Lantz Marion Salou Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies STAR Protocols Cell culture Immunology CRISPR |
| title | Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies |
| title_full | Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies |
| title_fullStr | Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies |
| title_full_unstemmed | Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies |
| title_short | Protocol to expand and CRISPR-Cas9 genomic edit murine MAIT cells for subsequent in vivo studies |
| title_sort | protocol to expand and crispr cas9 genomic edit murine mait cells for subsequent in vivo studies |
| topic | Cell culture Immunology CRISPR |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723003866 |
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