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Simonetti, M. ; Yilmazer, A.* ; Kretschmer, K.

Genetic tools for analyzing Foxp3+ treg cells: Fluorochrome-based transcriptional reporters and genetic fate-mapping.

Methods Mol. Biol. 2559, 95-114 (2023)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
The lack of unambiguous Foxp3+ Treg cell-specific surface markers has prompted the development of various transgenic mouse lines with Foxp3-dependent reporter activity, which involved different fluorochromes and transgenic strategies, including coexpression of multiple transgenes, such as Cre recombinase. Since then, Foxp3 transcriptional reporter has proven to be an indispensable tool to identify and isolate viable Foxp3+ Treg cell populations. However, the physiologic Treg cell pool is functionally heterogeneous and consists of intrathymically (tTreg) and peripherally (pTreg) induced Treg cells, which may confound interpretation of data relying on indiscriminatory Foxp3-fluorochrome reporter expressed in all Treg cells. In this chapter, we describe how the dual Foxp3RFP/GFP reporter can be exploited to discriminate both developmental sublineages based on tTreg cell lineage-specific GFP/Cre recombinase activity, in conjunction with Foxp3-driven RFP expression in all Foxp3+ Treg cells, and provide guidelines for experimental design and implementation. We also elaborate on the possibility to exploit GFP/Cre expression of Foxp3RFP/GFP reporter mice for the manipulation of gene expression (activation and inactivation), such as lineage tracing and in vivo ablation of tTreg cells, while sparing pTreg cells.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Cre-loxp ; Flow Cytometry ; Fluorochrome Reporter ; Foxp3 ; Transgenic Mice ; X-linked Inheritance ; Ttreg/ptreg
ISSN (print) / ISBN 1064-3745
e-ISSN 1940-6029
Book Volume Title Regulatory T-Cells
Journal Methods in Molecular Biology
Quellenangaben Volume: 2559, Issue: , Pages: 95-114 Article Number: , Supplement: ,
Publisher Springer
Publishing Place Berlin [u.a.]
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute for Pancreatic Beta Cell Research (IPI)
Grants Bundesministerium für Bildung und Forschung
Deutsche Forschungsgemeinschaft
German Center for Diabetes Research
FZT 111 (CRTD/Center for Regenerative Therapies Dresden