Download Project Title: Functional characterisation of centrosome proteins in

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.