Xist

Poster (download)

[pdf-embedder url=”https://bgrssb.icgbio.ru/wp-content/uploads/2020/07/165.pdf”]
Inna Pristyazhnyuk¹, Aleksei Menzorov^2
1Institute of Cytology and Genetics SB RAS, iprist@mail.ru
²Institute of Cytology and Genetics SB RAS, menzorov@bionet.nsc.ru

Induced pluripotent stem cells (iPSCs) were produced from many species. One of the key iPSC characteristics is X-chromosome inactivation status. Both active X-chromosomes mark naГЇve pluripotency, the presence of inactive X-chromosome suggests primed pluripotency. We had previously shown that American mink embryonic stem cells have both X-chromosomes active. In this study, we produced American mink iPSCs from XX embryonic fibroblasts and studied X-chromosome inactivation. We performed immunocytochemical staining of H3K27me3 chromatin modification (marker of inactive chromatin) and reverse transcription PCR of the Xist expression (marker of inactivated X-chromosome) in single cells. We have shown that according to the immunocytochemical analysis of three iPSC lines, from 56.8 to 95.6 % of cells had both X-chromosome homologs active. Xist expression data confirmed these results for one iPSC line, but unexpectedly the majority of cells in the other two iPSC lines were positive for Xist expression, so one of the X-chromosomes was presumably inactivated. Thus, we analyzed X-chromosome inactivation in XX American mink iPSCs for the first time and revealed the presence of different X-chromosome inactivation patterns.