Phosphoproteomic Analysis and Organotypic Cultures for the Study of Signaling Pathways
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Standard
Phosphoproteomic Analysis and Organotypic Cultures for the Study of Signaling Pathways. / Ye, Zilu; Wandall, Hans H.; Dabelsteen, Sally.
I: Bio-protocol, Bind 14, Nr. 4, e4941, 2024.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Phosphoproteomic Analysis and Organotypic Cultures for the Study of Signaling Pathways
AU - Ye, Zilu
AU - Wandall, Hans H.
AU - Dabelsteen, Sally
N1 - Publisher Copyright: © 2024 Bio-protocol LLC. All rights reserved.
PY - 2024
Y1 - 2024
N2 - Signaling pathways are involved in key cellular functions from embryonic development to pathological conditions, with a pivotal role in tissue homeostasis and transformation. Although most signaling pathways have been intensively examined, most studies have been carried out in murine models or simple cell culture. We describe the dissection of the TGF-β signaling pathway in human tissue using CRISPR-Cas9 genetically engineered human keratinocytes (N/TERT-1) in a 3D organotypic skin model combined with quantitative proteomics and phosphoproteomics mass spectrometry. The use of human 3D organotypic cultures and genetic engineering combined with quantitative proteomics and phosphoproteomics is a powerful tool providing insight into signaling pathways in a human setting. The methods are applicable to other gene targets and 3D cell and tissue models.
AB - Signaling pathways are involved in key cellular functions from embryonic development to pathological conditions, with a pivotal role in tissue homeostasis and transformation. Although most signaling pathways have been intensively examined, most studies have been carried out in murine models or simple cell culture. We describe the dissection of the TGF-β signaling pathway in human tissue using CRISPR-Cas9 genetically engineered human keratinocytes (N/TERT-1) in a 3D organotypic skin model combined with quantitative proteomics and phosphoproteomics mass spectrometry. The use of human 3D organotypic cultures and genetic engineering combined with quantitative proteomics and phosphoproteomics is a powerful tool providing insight into signaling pathways in a human setting. The methods are applicable to other gene targets and 3D cell and tissue models.
KW - Cell signaling
KW - Gene editing
KW - Keratinocytes
KW - Organotypic model
KW - Phosphoproteomics
KW - Proteomics
U2 - 10.21769/BioProtoc.4941
DO - 10.21769/BioProtoc.4941
M3 - Journal article
C2 - 38410375
AN - SCOPUS:85185558977
VL - 14
JO - Bio-protocol
JF - Bio-protocol
SN - 2331-8325
IS - 4
M1 - e4941
ER -
ID: 384618638