Functional Analysis of Foxp3 and Its Mutants by Retroviral Transduction of Murine Primary CD4+ T Cells

Akira Nakajima; Ryuichi Murakami; Shohei Hori

doi:10.1007/978-1-0716-2647-4_7

Functional Analysis of Foxp3 and Its Mutants by Retroviral Transduction of Murine Primary CD4⁺ T Cells

Methods Mol Biol. 2023:2559:79-94. doi: 10.1007/978-1-0716-2647-4_7.

Authors

Akira Nakajima¹, Ryuichi Murakami¹, Shohei Hori²

Affiliations

¹ Laboratory of Immunology and Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
² Laboratory of Immunology and Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan. shohei@mol.f.u-tokyo.ac.jp.

PMID: 36180628
DOI: 10.1007/978-1-0716-2647-4_7

Abstract

The transcription factor Foxp3/FOXP3 orchestrates regulatory T (Treg) cell development and function by interacting with numerous target genes and partner proteins. Functional analysis of naturally occurring or engineered Foxp3/FOXP3 mutations has provided important insights into how the complex Foxp3/FOXP3-centered molecular network operates. Here, we describe detailed protocols for retroviral transduction of murine primary conventional CD4⁺ T cells to determine the impacts of Foxp3 mutations on the Treg-cell-like phenotype and function conferred by Foxp3.

Keywords: Flow cytometry; Foxp3, mutagenesis; Regulatory T cells; Retroviral gene transduction.

Publication types

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

MeSH terms

Animals
CD4-Positive T-Lymphocytes* / metabolism
Forkhead Transcription Factors* / genetics
Forkhead Transcription Factors* / metabolism
Gene Expression Regulation
Mice
Retroviridae / genetics
Retroviridae / metabolism
T-Lymphocytes, Regulatory / metabolism

Substances

Forkhead Transcription Factors
Foxp3 protein, mouse