Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2023
Höfundar / Authors: Milan Pieter Paul de Putter, Halla Rós Ólafsdóttir, Kristján Hólm Grétarsson, Erna Magnúsdóttir
Starfsvettvangur / Affiliations: Erna Magnúsdóttir´s lab, Department of Anatomy and Biomedical Science, School of Health Sciences, University of Iceland
Kynnir / Presenter: Milan Pieter Paul de Putter
Primordial germ cells (PGCs) are the founders of the germ line and therefore essential for the transfer of genetic and epigenetic information from one generation to the next. It is therefore imperative to understand PGC specification and germ cell line development to tackle issues in reproductive biology, e.g. sub- or infertility. We aim to elucidate the functions of the RHOX5 and RHOX6 transcriptional regulators within mouse PGCs.
The Rhox genes form an X-linked cluster that are expressed colinearly in the germ cells. Furthermore, preliminary data link the RHOX factors to the expression of germ cell genes and repression of DNA methylation. We propose to use Rapid Immunoprecipitation and Mass Spectrometry of Endogenous proteins (RIME) to uncover protein interactors involved in RHOX5/6-mediated transcriptional regulation and gain insight into the cis-regulatory logic governing their function. Through subsequent knock-out of these co-binding factors, the relation to PGC specification can be investigated.
The project utilizes endogenous or doxycycline-inducible overexpression of HA- or 3xTY1-tagged RHOX5/6 mouse embryonic stem cells. These can be induced first into epiblast-like cells and then into PGC-like cells to yield three distinct developmental stages to analyze RHOX factor expression and regulation. This provides a strong framework to obtain fundamental knowledge of germ cell and homeobox factor biology with relevance for regenerative medicine and reproductive biology.