Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2017
Erindi/veggspjald / Talk/poster E62
Höfundar / Authors: Erna Magnúsdóttir, Kristján Hólm Grétarsson, Daisy Awiti
Starfsvettvangur / Affiliations: Læknadeild Háskóla Íslands, Vatnsmýrarvegi 16, 101 Reykjavík
Kynnir / Presenter: Erna Magnúsdóttir
Prior to implantation, the inner cell mass of a mouse embryo is said to have naive pluripotency that later progresses to primed pluripotency of the post-implantation epiblast. The cells of the epiblast can subsequently assume primordial germ cell fate exhibiting underlying pluripotency. Numerous factors have been identified as critical for these pluripotent state transitions. We are working on identifying transcriptional regulators that take part in these state transitions.
In order to investigate potential transcription factors regulating the transition from naive to primed pluripotency of mouse embryonic stem cells we have generated loss of function mutant cell lines using CRISPR-Cas9. The cell lines contain transcriptional reporters for naive pluripotency (GOF cells, containing GFP under the control of the distal enhancer and basal promoter of Pou5f1-Oct4) and primordial cerm cell fate. Using these systems we have identified a transcriptional regulator that is important for the exit from naive pluripotency facilitated by a switch from 2i/Lif media (containing inhibitors of GSK3beta and Fgf signalling), and as well as the induction of mouse primordial germ cells by BMP4. Lineage commitement factors as well as primordial germ cell markers are aberrantly expressed in the loss of function mutants. We are now testing lineage commitment dynamices of the mutants and assessing the transription factor DNA binding patterns.