Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
CRISPR-mediated endogenous tagging of genes provides unique possibilities to explore the function and dynamic subcellular localization of proteins in living cells. Here, we describe experimental strategies for endogenous PCR-tagging of ciliary genes in human RPE1 cells and how image acquisition and analysis of the expressed fluorescently tagged proteins can be utilized to study the dynamic ciliary processes of intraflagellar transport and vesicular trafficking.
| Original language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Number of pages | 20 |
| Publisher | Humana Press |
| Publication date | 2024 |
| Pages | 147-166 |
| Chapter | 9 |
| ISBN (Print) | 978-1-0716-3506-3 |
| ISBN (Electronic) | 978-1-0716-3507-0 |
| DOIs | |
| Publication status | Published - 2024 |
| Series | Methods in Molecular Biology |
|---|---|
| Volume | 2725 |
| ISSN | 1064-3745 |
Bibliographical note
Funding Information:
We thank Joel Pomerantz, Kazuhisa Nakayama, Michael Knop, Keith Joung, and Benjamin Kleinstiver for reagents and the Danish Molecular Biomedical Imaging Center, University of Southern Denmark, for use of imaging equipment, supported by Novo Nordisk Foundation (NNF18SA0032928). This work was supported by grants from Independent Research Fund Denmark (grant 8021-00425B) to J.S.A. and Novo Nordisk Foundation (NNF18OC0053024, NNF15OC0016886 and NNF14OC0011535), The Danish Cancer Society (R146-A9590) and Hartmann Foundation (A31662) to L.B.P.
Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.
- Cas12a, Cas9, Cilia, CRISPR, Fluorescently tagged proteins, Image analysis, Intraflagellar transport, Live-cell imaging, Vesicle trafficking
Research areas
ID: 373795328