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Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging
Stefanie Kuhns1, Alice Dupont Juhl1, Zeinab Anvarian2
1Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark.
Methods in Molecular Biology (Clifton, N.J.)
|October 19, 2023
Abstract:
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.
