University of Nottingham, School of Biosciences

Masaki Kinoshita

Formative state of pluripotency represents the early post-implantation stage epiblast cells and formative stem (FS) cells were established from such stage of the mouse embryos. It has unique features such as blastocyst chimaera competency and direct germ cell differentiation potency. In formative phase, pre-implantation stage naïve blastocyst cells become competent to respond to differentiation signals, which each cell can initiate the gastrulation. To understand the gene regulatory network (GRN) to support these unique features, we performed CRISPR/gRNA based screening to identify genes required for the self-renewal of FS cells. We successfully identified 1843 genes, which is essential for the proliferation. In this project, we will focus on several genes to perform the detailed molecular analyses. We will establish FKBP and HA tag knock-in FS cell lines. Using such cell lines, we will perform gene expression analysis at various time points after the induction of protein degradation by dTAG1. We will also perform Cut&Run experiments using HA-tag to identify their biding sites on the genome to understand formative GRN.