Projektdetails
Abstract
Development of higher eukaryotic organisms proceeds through a series of cell fate transitions, typically
accompanied by a decline in developmental potency. During embryogenesis all somatic tissues are derived from
the pluripotent cells of the inner cell mass. Research in recent years has contributed to a substantial
understanding of the molecular underpinnings of self-renewal and pluripotency. However, the factors that compute
cues from the cellular environment to elicit an integrated and exact series of cell-fate choices remain largely
elusive. Their identification and knowledge of their molecular role will be a crucial milestone in devising efficient
and deterministic protocols to drive lineage specification. Work proposed here will be a major milestone towards
this goal. To this end, we will combine genetic screens, detailed gene- and pathway-level analysis and systemsbiology
approaches to explore the mechanistic basis of the key cell fate decision that defines the exit from
pluripotency.
We have established haploid ES cells as a powerful screening platform to interrogate developmental processes
and have performed a saturation screen to identify central players involved in the exit from self-renewal. To
analyse screen data and to build a statistical framework for analysis of transposon screens, the Leeb (stem cell
genetics) and Beyer (computational systems biology) groups have established a standing collaboration. In this
proposal we request funding for two post-doctoral researchers, one technician and two Masters students to
embark on a highly collaborative and ambitious project to generate 200 isogenic and passage matches mutant
ES cell lines (for 100 Genes, two clones each), followed by detailed gene expression analysis in order to identify
the core regulatory mechanisms that drive the exit from naive pluripotency.
Here we propose to use high-throughput cell culture approaches combined with sophisticated and customized
computational tools to allow us to approach ES cell differentiation in a systems biology approach. Specifically,
one main aim of this project is to delineate the genetic switches that gate the exit from naïve pluripotency by
studying a series of genetic perturbation of differnetiation. The combination of expertise in the Beyer and Leeb
laboratories will position us ideally to identify relevant candidate genes, regulatory nodes and to delineate the
exact molecular mode of action of candidate-genes and pathways using controlled genetic and biochemical
experiments under defined culture conditions.
The work package laid out in this proposal is timely and highly relevant in our endeavour to generate a clear
understanding of the mechanisms that initiate ES cell differentiation, their central downstream targets and
potential genetic interactions. This will constitute a major contribution towards a better
accompanied by a decline in developmental potency. During embryogenesis all somatic tissues are derived from
the pluripotent cells of the inner cell mass. Research in recent years has contributed to a substantial
understanding of the molecular underpinnings of self-renewal and pluripotency. However, the factors that compute
cues from the cellular environment to elicit an integrated and exact series of cell-fate choices remain largely
elusive. Their identification and knowledge of their molecular role will be a crucial milestone in devising efficient
and deterministic protocols to drive lineage specification. Work proposed here will be a major milestone towards
this goal. To this end, we will combine genetic screens, detailed gene- and pathway-level analysis and systemsbiology
approaches to explore the mechanistic basis of the key cell fate decision that defines the exit from
pluripotency.
We have established haploid ES cells as a powerful screening platform to interrogate developmental processes
and have performed a saturation screen to identify central players involved in the exit from self-renewal. To
analyse screen data and to build a statistical framework for analysis of transposon screens, the Leeb (stem cell
genetics) and Beyer (computational systems biology) groups have established a standing collaboration. In this
proposal we request funding for two post-doctoral researchers, one technician and two Masters students to
embark on a highly collaborative and ambitious project to generate 200 isogenic and passage matches mutant
ES cell lines (for 100 Genes, two clones each), followed by detailed gene expression analysis in order to identify
the core regulatory mechanisms that drive the exit from naive pluripotency.
Here we propose to use high-throughput cell culture approaches combined with sophisticated and customized
computational tools to allow us to approach ES cell differentiation in a systems biology approach. Specifically,
one main aim of this project is to delineate the genetic switches that gate the exit from naïve pluripotency by
studying a series of genetic perturbation of differnetiation. The combination of expertise in the Beyer and Leeb
laboratories will position us ideally to identify relevant candidate genes, regulatory nodes and to delineate the
exact molecular mode of action of candidate-genes and pathways using controlled genetic and biochemical
experiments under defined culture conditions.
The work package laid out in this proposal is timely and highly relevant in our endeavour to generate a clear
understanding of the mechanisms that initiate ES cell differentiation, their central downstream targets and
potential genetic interactions. This will constitute a major contribution towards a better
Status | Abgeschlossen |
---|---|
Tatsächlicher Beginn/ -es Ende | 1/03/18 → 28/02/21 |
Projektbeteiligte
- Universität Wien (Leitung)
- Universität zu Köln