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

Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt

Standard

Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging. / Kuhns, Stefanie; Juhl, Alice Dupont; Anvarian, Zeinab; Wüstner, Daniel; Pedersen, Lotte B.; Andersen, Jens S.

Methods in Molecular Biology. Humana Press, 2024. s. 147-166 (Methods in Molecular Biology, Bind 2725).

Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt

Harvard

Kuhns, S, Juhl, AD, Anvarian, Z, Wüstner, D, Pedersen, LB & Andersen, JS 2024, Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging. i Methods in Molecular Biology. Humana Press, Methods in Molecular Biology, bind 2725, s. 147-166. https://doi.org/10.1007/978-1-0716-3507-0_9

APA

Kuhns, S., Juhl, A. D., Anvarian, Z., Wüstner, D., Pedersen, L. B., & Andersen, J. S. (2024). Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging. I Methods in Molecular Biology (s. 147-166). Humana Press. Methods in Molecular Biology Bind 2725 https://doi.org/10.1007/978-1-0716-3507-0_9

Vancouver

Kuhns S, Juhl AD, Anvarian Z, Wüstner D, Pedersen LB, Andersen JS. Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging. I Methods in Molecular Biology. Humana Press. 2024. s. 147-166. (Methods in Molecular Biology, Bind 2725). https://doi.org/10.1007/978-1-0716-3507-0_9

Author

Kuhns, Stefanie ; Juhl, Alice Dupont ; Anvarian, Zeinab ; Wüstner, Daniel ; Pedersen, Lotte B. ; Andersen, Jens S. / Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging. Methods in Molecular Biology. Humana Press, 2024. s. 147-166 (Methods in Molecular Biology, Bind 2725).

Bibtex

@inbook{a687b14762b242469c9b3d79fa617f19,
title = "Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging",
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.",
keywords = "Cas12a, Cas9, Cilia, CRISPR, Fluorescently tagged proteins, Image analysis, Intraflagellar transport, Live-cell imaging, Vesicle trafficking",
author = "Stefanie Kuhns and Juhl, {Alice Dupont} and Zeinab Anvarian and Daniel W{\"u}stner and Pedersen, {Lotte B.} and Andersen, {Jens S.}",
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: {\textcopyright} The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.",
year = "2024",
doi = "10.1007/978-1-0716-3507-0_9",
language = "English",
isbn = "978-1-0716-3506-3",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "147--166",
booktitle = "Methods in Molecular Biology",
address = "United States",

}

RIS

TY - CHAP

T1 - Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging

AU - Kuhns, Stefanie

AU - Juhl, Alice Dupont

AU - Anvarian, Zeinab

AU - Wüstner, Daniel

AU - Pedersen, Lotte B.

AU - Andersen, Jens S.

N1 - 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.

PY - 2024

Y1 - 2024

N2 - 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.

AB - 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.

KW - Cas12a

KW - Cas9

KW - Cilia

KW - CRISPR

KW - Fluorescently tagged proteins

KW - Image analysis

KW - Intraflagellar transport

KW - Live-cell imaging

KW - Vesicle trafficking

U2 - 10.1007/978-1-0716-3507-0_9

DO - 10.1007/978-1-0716-3507-0_9

M3 - Book chapter

C2 - 37856023

AN - SCOPUS:85175193335

SN - 978-1-0716-3506-3

T3 - Methods in Molecular Biology

SP - 147

EP - 166

BT - Methods in Molecular Biology

PB - Humana Press

ER -

ID: 373795328