A protocol for generation and live-cell imaging analysis of primary cilia reporter cell lines

Lucia Binó; Erika Mikulenková; Luděk Štepánek; Ondřej Bernatík; David Vysloužil; Petra Pejšková; Peter Gorilák; Martina Huranová; Vladimír Varga; Lukáš Čajánek

doi:10.1016/j.xpro.2022.101199

A protocol for generation and live-cell imaging analysis of primary cilia reporter cell lines

STAR Protoc. 2022 Mar 2;3(1):101199. doi: 10.1016/j.xpro.2022.101199. eCollection 2022 Mar 18.

Authors

Affiliations

¹ Laboratory of Cilia and Centrosome Biology, Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 3, 62500 Brno, Czechia.
² Laboratory of Cell Motility, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czechia.
³ Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5, 62500 Brno, Czechia.
⁴ Charles University, Faculty of Science, Albertov 6, 128 00 Prague, Czechia.
⁵ Laboratory of Adaptive Immunity, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czechia.

Abstract

Primary cilia are hair-like sensory organelles protruding from the surface of most human cells. As cilia are dynamic, several aspects of their biology can only be revealed by real-time analysis in living cells. Here we describe the generation of primary cilia reporter cell lines. Furthermore, we provide a detailed protocol of how to use the reporter cell lines for live-cell imaging microscopy analysis of primary cilia to study their growth as well as intraciliary transport. For complete details on the use and execution of this protocol, please refer to Bernatik et al. (2020) and Pejskova et al. (2020).

Keywords: Cell Biology; Cell culture; Microscopy; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cilia* / metabolism
Humans
Image Processing, Computer-Assisted* / methods
Microscopy / methods