Project Details
Projekt
Functional analysis of H3 methylation writers and erasers regulated at the end of neurogenesis
Applicant
Professorin Dr. Magdalena Götz
Subject Area
Developmental Neurobiology
Molecular Biology and Physiology of Neurons and Glial Cells
Molecular Biology and Physiology of Neurons and Glial Cells
Term
since 2025
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 563413066
Neurogenesis comes to a permanent end in most regions of the mammalian brain, which is one of the reasons why degenerated neurons after brain injury are not regenerated. Here we aim to explore the epigenetic factors contributing to ending neurogenesis with the ultimate goal to use them to activate neurogenesis in glial cells. To identify novel candidates, we performed a multiome profiling of neurogenic versus gliogenic cortical radial glial cells, the neural stem cells, isolated at E14, the peak of neurogenesis, and E18, the end of neurogenesis and transition to gliogenesis, respectively. This showed a profound down-regulation of a network of histone H3 methyltransferases and demethylases, which was further corroborated by analysis of RNA-seq data from human fetal cortex comparing stages during neurogenesis and gliogenesis. Here we therefore aim to explore the function of these enzymes in the murine cortex in vivo and human iPSC-derived neuronal and glial progenitors in vitro and to probe the potential of histone H3 methylation manipulation in promoting maintenance of neurogenic gene expression at the time when neural stem cells are normally progressing to gliogenesis. In Aim 1 we will perform a small targeted CRISPR-knock-out screen to determine the function of these candidate enzymes in neurogenesis, including characterization of the respective histone marks targeted by the functionally relevant enzymes. In Aim2 we plan to probe if these enzymes are sufficient to activate neurogenic gene expression at the onset of gliogenesis in the murine cerebral cortex using CRISPRa, followed in Aim 3 by epigenetic engineering using the respective enzymes fused to dCas9 targeting the neurogenic genes identified in Aims 1 and 2. In Aim 4 we will explore the function of these enzymes in human iPSC-derived neuronal and glial progenitors first mapping the main differences in these marks between these cell types using Epi-CYTOF and then activating the respective enzymes in the glial progenitors to examine gene expression changes. These approaches will allow understanding the role of H3 methylation in neurogenesis and how the loss of several writers and erasers contributes to the switch to gliogenesis.
DFG Programme
Priority Programmes
