Ciliary Assembly/Disassembly Assay in Non-transformed Cell Lines
Masaki Saito,
Kensuke Sakaji,
Wataru Otsu,
Ching-Hwa Sung
Affiliations
Masaki Saito
Department of Molecular Pharmacology, Tohoku University School of Medicine, Sendai, Japan
Kensuke Sakaji
Department of Molecular Pharmacology, Tohoku University School of Medicine, Sendai, Japan
Wataru Otsu
Department of Ophthalmology, Margaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY, USA
Ching-Hwa Sung
Department of Ophthalmology, Margaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY, USADepartment of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY, USA, Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan
The primary cilium is a non-motile sensory organelle whose assembly and disassembly are closely associated with cell cycle progression. The primary cilium is elongated from the basal body in quiescent cells and is resorbed as the cells re-enter the cell cycle. Dysregulation of ciliary dynamics has been linked with ciliopathies and other human diseases. The in vitro serum-stimulated ciliary assembly/disassembly assay has gained popularity in addressing the functions of the protein-of-interest in ciliary dynamics. Here, we describe a well-tested protocol for transfecting human retinal pigment epithelial cells (RPE-1) and performing ciliary assembly/disassembly assays on the transfected cells.
