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Ciliary Assembly/Disassembly Assay in Non-transformed Cell Lines.
Masaki Saito1, Kensuke Sakaji1, Wataru Otsu2
1Department of Molecular Pharmacology, Tohoku University School of Medicine, Sendai, Japan.
Bio-Protocol
|June 28, 2021
Summary
This study details a protocol for studying primary cilia dynamics in RPE-1 cells. The method facilitates research into ciliopathies and human diseases linked to ciliary dysfunction.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- The primary cilium is a crucial sensory organelle involved in cell cycle regulation.
- Aberrant ciliary assembly and disassembly are implicated in ciliopathies and other diseases.
- Understanding ciliary dynamics is vital for disease research.
Purpose of the Study:
- To present a reliable protocol for assessing primary cilium assembly and disassembly.
- To enable the study of protein functions in ciliary dynamics using transfected RPE-1 cells.
Main Methods:
- Development and validation of a serum-stimulated ciliary assembly/disassembly assay.
- Protocol for transfecting human retinal pigment epithelial (RPE-1) cells.
- Performing *in vitro* ciliary assays on transfected cells.
Main Results:
- A well-tested protocol for RPE-1 cell transfection is established.
- The assay allows for the investigation of ciliary dynamics in response to serum stimulation.
- This method supports the functional analysis of proteins involved in ciliary assembly/disassembly.
Conclusions:
- The described protocol provides a robust method for studying primary cilium dynamics.
- This assay is valuable for investigating the molecular mechanisms underlying ciliopathies and related diseases.
- The protocol facilitates the functional characterization of proteins impacting ciliary behavior.
