1. Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis.
- Author
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Falk, Nathalie, Kessler, Kristin, Schramm, Sinja-Fee, Boldt, Karsten, Becirovic, Elvir, Michalakis, Stylianos, Regus-Leidig, Hanna, Noegel, Angelika A., Ueffing, Marius, Thiel, Christian T., Roepman, Ronald, Brandstätter, Johann Helmut, and Gießl, Andreas
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RETINA , *CRISPRS , *CILIOPATHY - Abstract
Pericentrin (Pcnt) is a multifunctional scaffold protein and mutations in the human PCNT gene are associated with several diseases, including ciliopathies. Pcnt plays a crucial role in ciliary development in olfactory receptor neurons, but its function in the photoreceptorconnecting cilium is unknown. We downregulated Pcnt in the retina ex vivo and in vivo via a virus-based RNA interference approach to study Pcnt function in photoreceptors. ShRNA-mediated knockdown of Pcnt impaired the development of the connecting cilium and the outer segment of photoreceptors, and caused a nuclear migration defect. In protein interaction screens, we found that the outer nuclear membrane protein Syne-2 (also known as Nesprin-2) is an interaction partner of Pcnt in photoreceptors. Syne-2 is important for positioning murine photoreceptor cell nuclei and for centrosomal migration during early ciliogenesis. CRISPR/Cas9-mediated knockout of Syne-2 in cell culture led to an overexpression and mislocalization of Pcnt and to ciliogenesis defects. Our findings suggest that the Pcnt-Syne-2 complex is important for ciliogenesis and outer segment formation during retinal development and plays a role in nuclear migration. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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