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Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice
Author(s) -
Fraser Stuart T.,
Hadjantonakis AnnaKaterina,
Sahr Kenneth E.,
Willey Stephen,
Kelly Olivia G.,
Jones Elizabeth A.V.,
Dickinson Mary E.,
Baron Margaret H.
Publication year - 2005
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.20139
Subject(s) - biology , histone h2b , microbiology and biotechnology , yellow fluorescent protein , fusion protein , transgene , chromatin , histone , embryonic stem cell , green fluorescent protein , genetics , gene , recombinant dna
Abstract We report the first endothelial lineage‐specific transgenic mouse allowing live imaging at subcellular resolution. We generated an H2B‐EYFP fusion protein which can be used for fluorescent labeling of nucleosomes and used it to specifically label endothelial cells in mice and in differentiating embryonic stem (ES) cells. A fusion cDNA encoding a human histone H2B tagged at its C‐terminus with enhanced yellow fluorescent protein (EYFP) was expressed under the control of an Flk1 promoter and intronic enhancer. The Flk1::H2B‐EYFP transgenic mice are viable and high levels of chromatin‐localized reporter expression are maintained in endothelial cells of developing embryos and in adult animals upon breeding. The onset of fluorescence in differentiating ES cells and in embryos corresponds with the beginning of endothelial cell specification. These transgenic lines permit real‐time imaging in normal and pathological vasculogenesis and angiogenesis to track individual cells and mitotic events at a level of detail that is unprecedented in the mouse. genesis 42:162–171, 2005. © 2005 Wiley‐Liss, Inc.