BLIMP1 IN PGCS · Molecular Mechanism for Primordial Germ Cell Specification - The Role of Blimp1
FP7 — People (Marie Curie Actions)
- Duration
- 2009-06-01 → 2011-05-31
- EU contribution
- €181,351
- Participants
- 1
- Scheme
- MC-IEF
Lines connect the coordinator with its partners.
Results in brief
Molecular mechanism for primordial germ cell specification - the role of Blimp1
Germ cells serve the critical function of transmitting genetic and epigenetic information across generations in multi-cellular organisms. A unique role of the germ line is to generate the totipotent state. Furthermore, while the early primordial germ cells (PGCs) exhibit many features of pluripotency, this lineage is functionally unipotent. Studies of this cell lineage are potentially of wide general impact in the context of stem cells and mechanisms underlying pluripotency. The purpose of the current project is to address the molecular mechanism of events that accompany specification of the germ cells, which distinguishes them from the neighbouring somatic cells that share common ancestry. The focus of my work has been the transcription factor Blimp1, with the main objective of uncovering the precise molecular mechanism of its action in PGC specification. To this end, I generated genome wide DNA binding profiles for Blimp1 in the context of pluripotent cells. I have also performed gene expression profiling to assess the transcriptional changes occurring downstream of Blimp1 in pluripotent cells. Integrating the data sets reveals primary and secondary events occurring downstream of Blimp1. I have also compared the DNA binding profiles to published results for other factors important for pluripotency, revealing potential mechanisms for Blimp1 acting in conjunction with other factors in PCG specification.
Data: CORDIS, © European Union
Project objective
Germ cells serve the critical function of transmitting genetic and epigenetic information across generations in multi-cellular organisms. An important attribute of the germ line is to generate the totipotent state. Studies of this unique cell lineage are of wide general interest in the context of stem cells and mechanisms of pluripotency. Relatively little is yet known about how germ cells are specified in mice. Recent genetic studies showed that the transcriptional repressor Blimp1 is a critical determinant of germ cell fate in the epiblast cells of the mouse embryo. There are only about 35 founder primordial germ cells (PGCs) present in each embryo, providing a significant hurdle for the mechanistic elucidation of germ cell specification. A robust in-vitro system that mimics PGC specification is essential to gain a deeper insight into the role of Blimp1 in germ cell fate determination. This project aims to generate such a system using a twofold approach. First, by engineering embryonic stem cells to express Blimp1 in an inducible manner and utilizing them for the induction of PGC fate. Second, by generating transgenic mice expressing fluorescent reporter molecules, for the derivation of epiblast stem cell cultures and subsequent FACS sorting of PGCs. To elucidate the gene expression programme governed by Blimp1 during PGC specification, the culture models will be used for analysis of the genome wide target site binding of Blimp1 and performing functional assays of the target genes and co-repressor proteins bound with Blimp1 to the sites identified. This will likely provide novel biochemical insight into PGC specification revealing mechanisms underlying the balance between pluripotency and differentiation. The project will provide training to the fellow in terms of techniques and subjects pertaining to stem cells, germ cells, mouse genetics and development, while at the same time allowing the re-integration of the fellow into the European scientific community.
Original text from CORDIS.
Participants
- THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGECoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union