MHC II-EGFP Knock-in Mouse Model

Jan Pačes, Valéria Grobárová, Zdeněk Zadražil, Karolina Knížková, Nikola Malinská, Liliana Tušková, Marianne Boes, Jan Černý*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

The MHC II–EGFP knock-in mouse model enables us to visualize and track MHC-II-expressing cells in vivo by expressing enhanced green fluorescent protein (EGFP) fused to the MHC class II molecule under the MHC II beta chain promoter. Using this model, we can easily identify MHC-II-expressing cells, including dendritic cells, B cells, macrophages, and ILC3s, which play a key role as antigen-presenting cells (APCs) for CD4+ T cells. In addition, we can also precisely identify and analyze APC-containing tissues and organs. Even after fixation, EGFP retains its fluorescence, so this model is suitable for immunofluorescence studies, facilitating an unbiased characterization of the histological context, especially with techniques such as light-sheet fluorescence microscopy. Furthermore, the MHC II–EGFP knock-in mouse model is valuable for studying the molecular mechanisms of MHC II gene regulation and expression by making it possible to correlate MHC II expression (MHC II–EGFP) with surface fraction through antibody detection, thereby shedding light on the intricate regulation of MHC II expression.

Original languageEnglish
Article numbere925
JournalCurrent Protocols
Volume3
Issue number11
DOIs
Publication statusPublished - Nov 2023

Keywords

  • Animals
  • Antigen-Presenting Cells
  • B-Lymphocytes
  • Cell Membrane
  • Disease Models, Animal
  • Green Fluorescent Proteins
  • Mice

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