| Verfasst von: | Ambrosi, Giulia [VerfasserIn] |
|---|---|
| Voloshanenko, Oksana [VerfasserIn] | |
| Eckert, Antonia [VerfasserIn] | |
| Kranz, Dominique [VerfasserIn] | |
| Nienhaus, G. Ulrich [VerfasserIn] | |
| Boutros, Michael [VerfasserIn] | |
| Titel: | Allele-specific endogenous tagging and quantitative analysis of β-catenin in colorectal cancer cells |
| Verf.angabe: | Giulia Ambrosi, Oksana Voloshanenko, Antonia F. Eckert, Dominique Kranz, G. Ulrich Nienhaus, Michael Boutros |
| E-Jahr: | 2022 |
| Jahr: | 11 January 2022 |
| Umfang: | 27 S. |
| Fussnoten: | Gesehen am 15.02.2022 |
| Titel Quelle: | Enthalten in: eLife |
| Ort Quelle: | Cambridge : eLife Sciences Publications, 2012 |
| Jahr Quelle: | 2022 |
| Band/Heft Quelle: | 11(2022), Artikel-ID e64498, Seite 1-27 |
| ISSN Quelle: | 2050-084X |
| Abstract: | Wnt signaling plays important roles in development, homeostasis, and tumorigenesis. Mutations in beta-catenin that activate Wnt signaling have been found in colorectal and hepatocellular carcinomas. However, the dynamics of wild-type and mutant forms of beta-catenin are not fully understood. Here, we genome-engineered fluorescently tagged alleles of endogenous beta-catenin in a colorectal cancer cell line. Wild-type and oncogenic mutant alleles were tagged with different fluorescent proteins, enabling the analysis of both variants in the same cell. We analyzed the properties of both beta-catenin alleles using immunoprecipitation, immunofluorescence, and fluorescence correlation spectroscopy approaches, revealing distinctly different biophysical properties. In addition, activation of Wnt signaling by treatment with a GSK3 beta inhibitor or a truncating APC mutation modulated the wild-type allele to mimic the properties of the mutant beta-catenin allele. The one-step tagging strategy demonstrates how genome engineering can be employed for the parallel functional analysis of different genetic variants. |
| DOI: | doi:10.7554/eLife.64498 |
| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt. Volltext: https://doi.org/10.7554/eLife.64498 |
| DOI: https://doi.org/10.7554/eLife.64498 | |
| Datenträger: | Online-Ressource |
| Sprache: | eng |
| Sach-SW: | adhesion |
| armadillo | |
| beta-catenin | |
| binding | |
| Cancer | |
| complex | |
| conversion | |
| crispr | |
| ctnnb1 | |
| efficient | |
| endogenous tagging | |
| fcs | |
| fluorescence correlation spectroscopy | |
| Human | |
| Oncogenic signaling | |
| plasma-membrane | |
| proteins | |
| region | |
| reveals | |
| Wnt signaling | |
| K10plus-PPN: | 1789606454 |
| Verknüpfungen: | → Zeitschrift |