Enderle, Leonie. Exploring the developmental functions of fat cadherins in Drosophila and mammals. 2016, PhD Thesis, University of Basel, Faculty of Science.
Restricted to Repository staff only until 1 July 2018.
Official URL: http://edoc.unibas.ch/diss/DissB_11787
The size and transmembrane nature of the mammalian Fat cadherin Fat4 had previously hindered the use of biochemical approaches to gain insight into its molecular functions. Here I developed several cell culture tools that allowed me to overcome some of these limitations and to study Fat4 localization and function. I performed proximity-dependent biotin identification (BioID) and identified an array of potential novel Fat4 interactors that will serve as a useful resource for future studies. Beside a variety of developmental defects, Fat4 mutant mice exhibit prenatal renal cysts with regions of abnormal primary cilia. Therefore, to understand if Fat4 regulates primary cilia, as has been suggested for several PCP proteins, I tested the effect of Fat4 depletion on cilia formation and maintenance in cell culture. Dramatic effects on cilia maintenance and centrosome positioning and coherence were found by knockdown with several independent siRNAs. However, CRISPR/Cas9-mediated Fat4 knockout did not confirm a requirement of Fat4 in these processes and indicated that cilia and centrosome defects were likely RNAi off-target effects. This highlights the potential pitfalls of RNAi and should be regarded as a cautionary tale.
|Advisors:||McNeill, Helen and Affolter, Markus and Cabernard, Clemens|
|Faculties and Departments:||05 Faculty of Science > Departement Biozentrum > Growth & Development > Cell Biology (Affolter)|
|Bibsysno:||Link to catalogue|
|Number of Pages:||1 Online-Ressource (261 Seiten)|
|Last Modified:||26 Sep 2016 12:57|
|Deposited On:||26 Sep 2016 12:56|
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