Protocol for multi-modal single-cell RNA sequencing on M. tuberculosis-infected mouse lungs.
Bioinformatics
Cell Biology
Flow Cytometry/Mass Cytometry
Genomics
Immunology
Microbiology
Sequencing
Single Cell
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
17 03 2023
17 03 2023
Historique:
received:
21
10
2022
revised:
20
12
2022
accepted:
20
01
2023
medline:
23
10
2023
pubmed:
1
3
2023
entrez:
28
2
2023
Statut:
ppublish
Résumé
To elucidate how different immune cells contribute to control or progression of M. tuberculosis (Mtb) infection, we developed a technique to perform multi-modal single-cell RNA sequencing (scRNA-seq) from in vivo Mtb-infected lung macrophages. This protocol simultaneously acquires the transcriptome, surface marker expression, and bacterial phenotype of each infected cell. We describe steps for sorting Mtb-infected cells and staining with CITE-seq antibodies, as well as for methanol fixation and generation of scRNA-seq libraries. This protocol can be used on tissues derived from murine, nonhuman primate, and human infections. For complete details on the use and execution of this protocol, please refer to Pisu et al. (2021).
Identifiants
pubmed: 36853694
pii: S2666-1667(23)00060-6
doi: 10.1016/j.xpro.2023.102102
pmc: PMC9937979
pii:
doi:
Substances chimiques
Antibodies
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
102102Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.