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Project Title: Functional characterisation of centrosome proteins in stem cells Project leader: Gislene Pereira Project Description: Many stem cells divide asymmetrically thereby generating mother and daughter cells with distinct cell fates. The differential distribution of cell fate determinants and/or cellular components depends on the correct orientation of the mitotic spindle accordingly to the cell polarity axis. The centrosome plays a critical role in this process. As the major microtubuleorganising centre of animal cells, centrosomes contribute to mitotic spindle formation and positioning. In addition, centrosomes are required for the biogenesis of the primary cilium, which is a chemosensory organelle present in many stem cells. The molecular function of cilia in stem cells remains largely unexplored but is most likely related to modulation of signalling pathways, such as Wnt, Sonic Hedgehog or Notch. The aim of this project is to investigate the role of centrosomal proteins in asymmetric cell division of stem cells. For this, the successful candidate will employ mouse neural stem cells that can be induced to divide asymmetrically in vitro. By using a combination of advanced microscopy, proteomic approaches, CRISPR-Cas9 genome editing and RNA interference, she/he will investigate protein localization, complex formation and phenotypes associated with loss- and gain-of-function mutants in stem cells. These studies will be complemented by in vivo analysis using mouse models. References: Lerit, DA et al. (2013). Organelle asymmetry for proper fitness, function and fate. Chromosome Res. 3:271-286. Kuhns, S et al. (2013). The microtubule affinity regulating kinase MARK4 promotes axoneme extension during early ciliogenesis. J. Cell Biol., 200:505-522. Schmidt, KN et al. (2012). Cep164 mediates vesicular docking to the mother centriole during early steps of ciliogenesis, J. Cell Biol., 199:1083-1101. Methods that will be used: Cell culture, advanced light microscopy (direct and indirect fluorescence microscopy, super resolution microscopy, live cell imaging), electron microscopy, proteomic approaches (purification of protein complexes, co-immunoprecipitation), genome editing (CRISPR-Cas9, Zinc finger-nucleases) in addition to standard molecular and cell biology methods (cloning, FACS analysis). Collaboration Partners: The PhD candidate will be part of the collaborative research consortium SFB873 (“Maintenance and Differentiation of Stem Cells in Development and Disease”) of the University of Heidelberg and as such she/he will participate in joint seminars/retreats together with researchers working in different areas of stem cell biology. In addition, the candidate will work in close collaboration with following research groups: Prof. Dr. Mike Heinemann (Bioquant, HD) – super resolution microscopy. Prof. Dr. Ana Martin-Villalba (DKFZ – HD) - in vivo functional studies. Profile of candidate’s qualification: Applicants should have a degree in Biological Sciences with a sound background in cell and molecular biology, and special interest in research related to stem cells, asymmetric cell division, centrosomes/cilia function. Applicants should be highly motivated, self-driven and able to work independently and as part of a team of international researchers. Applications should be submitted via the HBIGS webpage (http://www.hbigs.uniheidelberg.de/). Informal enquiries can be sent directly to Gislene Pereira ([email protected]). Keywords: Stem cells, asymmetric cell division, centrosome, cilia, signalling pathways, neural development.