RT Journal Article
SR Electronic
T1 Visualizing histone H4K20me1 in knock-in mice expressing the mCherry-tagged modification-specific intracellular antibody
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2024.03.13.584715
DO 10.1101/2024.03.13.584715
A1 Sato, Yuko
A1 Takenoshita, Maoko
A1 Ueoka, Miku
A1 Ueda, Jun
A1 Yamagata, Kazuo
A1 Kimura, Hiroshi
YR 2024
UL http://biorxiv.org/content/early/2024/03/14/2024.03.13.584715.abstract
AB During development and differentiation, histone modifications dynamically change locally and globally, associated with transcriptional regulation, DNA replication and repair, and chromosome condensation. The level of histone H4 Lys20 monomethylation (H4K20me1) increases during the G2 to M phases of the cell cycle and is enriched in facultative heterochromatin, such as inactive X chromosomes in cycling cells. To track the dynamic changes of H4K20me1 in living cells, we have developed a genetically encoded modification-specific intracellular antibody (mintbody) probe that specifically binds to the modification. Here, we report the generation of knock-in mice in which the coding sequence of the mCherry-tagged version of the H4K20me1-mintbody is inserted into the Rosa26 locus. The knock-in mice, which ubiquitously expressed the H4K20me1-mintbody, developed normally and were fertile, indicating that the expression of the probe does not disturb the cell growth, development, or differentiation. Various tissues isolated from the knock-in mice exhibited nuclear fluorescence without the need for fixation. The H4K20me1-mintbody was enriched in inactive X-chromosomes in developing embryos and in XY bodies during spermatogenesis. The knock-in mice will be useful for the histochemical analysis of H4K20me1 in any cell types.Competing Interest StatementThe authors have declared no competing interest.