| |  Online-Ressource |
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| Verfasst von: | Ambrosi, Giulia [VerfasserIn]  |
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| | Voloshanenko, Oksana [VerfasserIn] |
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| | Eckert, Antonia [VerfasserIn] |
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| | Kranz, Dominique [VerfasserIn] |
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| | Nienhaus, G. Ulrich [VerfasserIn] |
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| | Boutros, Michael [VerfasserIn] |
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| Titel: | Allele-specific endogenous tagging and quantitative analysis of β-catenin in colorectal cancer cells |
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| Verf.angabe: | Giulia Ambrosi, Oksana Voloshanenko, Antonia F. Eckert, Dominique Kranz, G. Ulrich Nienhaus, Michael Boutros |
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| E-Jahr: | 2022 |
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| Jahr: | 11 January 2022 |
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| Umfang: | 27 S. |
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| Fussnoten: | Gesehen am 15.02.2022 |
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| Titel Quelle: | Enthalten in: eLife |
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| Ort Quelle: | Cambridge : eLife Sciences Publications, 2012 |
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| Jahr Quelle: | 2022 |
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| Band/Heft Quelle: | 11(2022), Artikel-ID e64498, Seite 1-27 |
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| ISSN Quelle: | 2050-084X |
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| 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. |
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| DOI: | doi:10.7554/eLife.64498 |
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| 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 |
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| | DOI: https://doi.org/10.7554/eLife.64498 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | adhesion |
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| | armadillo |
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| | beta-catenin |
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| | binding |
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| | Cancer |
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| | complex |
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| | conversion |
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| | crispr |
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| | ctnnb1 |
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| | efficient |
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| | endogenous tagging |
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| | fcs |
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| | fluorescence correlation spectroscopy |
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| | Human |
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| | Oncogenic signaling |
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| | plasma-membrane |
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| | proteins |
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| | region |
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| | reveals |
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| | Wnt signaling |
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| K10plus-PPN: | 1789606454 |
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| Verknüpfungen: | → Zeitschrift |
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Allele-specific endogenous tagging and quantitative analysis of β-catenin in colorectal cancer cells / Ambrosi, Giulia [VerfasserIn]; 11 January 2022 (Online-Ressource)
