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Translatome Profiling of Tissue-Resident Macrophages Using the RiboTag Approach.

Authors :
Kim JS
Haimon Z
Boura-Halfon S
Jung S
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2713, pp. 253-267.
Publication Year :
2024

Abstract

Global gene expression profiling has provided valuable insights into the specific contributions of different cell types to various physiological processes. Notably though, both bulk and single-cell transcriptomics require the prior retrieval of the cells from their tissue context to be analyzed. Isolation protocols for tissue macrophages are, however, notoriously inefficient and, moreover, prone to introduce considerable bias and artifacts. Here, we will discuss a valuable alternative, originally introduced by Amieux and colleagues. This so-called RiboTag approach allows, in combination with respective macrophage-specific Cre transgenic lines, to retrieve macrophage translatomes from crude tissue extracts. We will review our experience with this ingenious method, focusing on the study of brain macrophages, including microglia and border-associated cells. We will elaborate on the advantages of the RiboTag approach that render it a valuable complement to standard cell sorting-based profiling strategies, especially for the investigation of tissue macrophages.<br /> (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Details

Language :
English
ISSN :
1940-6029
Volume :
2713
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
37639128
Full Text :
https://doi.org/10.1007/978-1-0716-3437-0_17