An innovative approach to functionally study the effect of epigenetic modifications on stem cells, neural development and brain tumours.

• Applicant: Professor Dr. Stefan Stricker
• Subject Area: Molecular Biology and Physiology of Neurons and Glial Cells ; Developmental Neurobiology
• Term: from 2014 to 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 257151303

• Final Report Year: 2020

Final Report Abstract

The aim of the project was to investigate which role epigenetic processes play in the development of the nervous system and its diseases. Using a transgenic approach, we intended to analyse the immediate effects of loss of histone modifications through overexpression of histone mutants in mouse neural stem cells. Early in the implementation of the project, we deviated from the original experimental strategy due to emerging publications and budget restraints. Instead, we explored the options novel CRISPR approaches can provide for the investigation of epigenetic processes. In the context of the project, we have developed tools and strategies for epigenome editing and transcriptional engineering in the nervous system, developed tools and strategies for epigenomic screens and for gRNA multiplexing. Furthermore, we have introduced these experimental strategies (e.g. epigenome editing and transcriptional engineering) to a larger scientific audience. In summary, this project provides a solid base for the use of non-genetic CRISPR approaches for the investigation of epigenetic processes during development and disease of the nervous system.

Publications

• (2015). Brave new epigenomes: the dawn of epigenetic engineering. Genome Med 7, 59
  Köferle, A., Stricker, S.H., and Beck, S.
  (See online at https://doi.org/10.1186/s13073-015-0185-8)
• (2016). CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening. BMC Genomics 17, 917
  Koferle, A., Worf, K., Breunig, C., Baumann, V., Herrero, J., Wiesbeck, M., Hutter, L.H., Gotz, M., Fuchs, C., Beck, S., et al.
  (See online at https://doi.org/10.1186/s12864-016-3268-z)
• (2017). A Universal Protocol for Large-scale gRNA Library Production from any DNA Source. JoVE, e56264
  Koferle, A., and Stricker, S.H.
  (See online at https://doi.org/10.3791/56264)
• (2017). From profiles to function in epigenomics. Nat Rev Genet 18, 51-66
  Stricker, S.H., Koferle, A., and Beck, S.
  (See online at https://doi.org/10.1038/nrg.2016.138)
• (2018). A Customizable Protocol for String Assembly gRNA Cloning (STAgR). JoVE, e58556
  Breunig, C.T., Neuner, A.M., Giehrl-Schwab, J., Wurst, W., Götz, M., and Stricker, S.H.
  (See online at https://doi.org/10.3791/58556)
• (2018). DNA-Methylation: Master or Slave of Neural Fate Decisions? Front Neurosci 12, 5
  Stricker, S.H., and Gotz, M.
  (See online at https://doi.org/10.3389/fnins.2018.00005)
• (2018). One step generation of customizable gRNA vectors for multiplex CRISPR approaches through string assembly gRNA cloning (STAgR). PLoS One 13, e0196015
  Breunig, C.T., Durovic, T., Neuner, A.M., Baumann, V., Wiesbeck, M.F., Koferle, A., Gotz, M., Ninkovic, J., and Stricker, S.H.
  (See online at https://doi.org/10.1371/journal.pone.0196015)