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PhD position (Epigenetics & Gene Regulation): Lineage Reprogramming of Glia into Neurons

In the field of “Lineage Reprogramming of Glia into Neurons”, the research group of Professor Benedikt Berninger offers the following PhD projects:

Activity-dependent gene regulation in induced neurons

In the first part of the project we will culture astroglia from the postnatal cortex and transduce these with retroviruses encoding for Neurog2 or Ascl1/Dlx2 for conversion into induced glutamatergic and GABAergic neurons (Heinrich et al., 2011). After different times of differentiation, neurons will be exposed to treatments that alter neural activity: high K+ (40mM), glutamate receptor agonists, or electrical field stimulation using frequencies known to induce long-term plasticity. Subsequently, induced neurons will be harvested for FACS-purification and RNA-sequencing. Using bioinformatics analyses in collaboration with Dr. Tiwari (IMB), we will identify genes expressed differentially upon stimulation. These gene expression changes will be compared to those of primary neurons undergoing the same stimulations. This will allow us to identify potential differences in the ability of induced and endogenous neurons to adapt their gene expression programs. We hypothesize that inability to fully unfold an activity-dependent gene regulation maybe due to failure to erase epigenetic modifications that were placed earlier during gliogenesis. Identifying such loci will be of great importance to characterize epigenetic barriers of lineage conversion into fully functional neurons.  
In the second part of the project we will investigate whether neurons derived from lineage converted glia in vivo have the ability to respond to network activity. Toward this aim, we will induce glia-to-neuron conversion by forced expression of Sox/Ascl1 (Heinrich et al., 2014) or Neurog2/Bcl2 (Gascón et al., 2016) in the cerebral cortex of adult mice following a mild stab wound injury. Concomitant with the reprogramming factors we will express Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) selectively either in induced neurons or the surrounding neuronal network. After allowing induced neurons to mature in vivo, we will activate DREADDs by their cognate ligand (CNO). We will then prepare brain slices to record from induced neurons (Heinrich et al., 2014) or endogenous neurons and collect cytoplasm for library preparation and RNA-sequencing (patch-seq) to compare alterations of the gene expression program between induced and endogenous neurons. The degree to which induced neurons are able to unfold an activity-dependent gene expression program will reveal to which extent these cells share the capacity for adaptive changes. Loci that fail to be activated in induced neurons will be compared to those identified in the in vitro part of the project. In a follow-up project we will then subject these loci to extensive analyses of their epigenetic state following gliogenesis and subsequent glia-to-neuron lineage conversion.             

We offer
  • The possibility to work on a cutting-edge project using state-of-the-art technology in a highly motivated research team
  • A stimulating, diverse and international research environment
  • Advanced training opportunities
  • A fully funded PhD position
Required qualifications
  • Master or Diploma
  • Motivation to solve complex biological problems
  • Excellent communication skills
Further requirement
  • 2 letters of reference
Duration of stipend/salary: 3 years, with the possibility of extension
Starting date: 1 August 2017 or later
Deadline for registration (exclusively online via web form): 24 March 2017

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While applying for the job please refer to jobvector and use the following reference number: JV-C12-BB

About Institute of Molecular Biology gGmbH

Our International PhD Programme on the “Gene Regulation, Epigenetics and DNA Damage Response” gives talented and enthusiastic students the opportunity to undertake PhD research at the cutting edge of modern biology. Our groups cover a broad range of expertise and include leading biochemists, geneticists, cell and developmental biologists who study the molecular mechanisms of...

More about Institute of Molecular Biology gGmbH

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