Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging

Stefanie Kuhns, Alice Dupont Juhl, Zeinab Anvarian, Daniel Wüstner, Lotte B. Pedersen, Jens S. Andersen*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-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 languageEnglish
Title of host publicationCilia : Methods and Protocols
EditorsVito Mennella
PublisherHumana Press
Publication date2024
ISBN (Print)978-1-0716-3509-4, 978-1-0716-3506-3
ISBN (Electronic)978-1-0716-3507-0
Publication statusPublished - 2024
SeriesMethods in Molecular Biology


  • Cas12a
  • Cas9
  • Cilia
  • Fluorescently tagged proteins
  • Image analysis
  • Intraflagellar transport
  • Live-cell imaging
  • Vesicle trafficking
  • Cilia/metabolism
  • Proteins/metabolism
  • Humans
  • Biological Transport
  • Protein Transport/genetics


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