| |  Online-Ressource |
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| Verfasst von: | Baldering, Tim N. [VerfasserIn]  |
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| | Karathanasis, Christos [VerfasserIn] |
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| | Harwardt, Marie-Lena I. E. [VerfasserIn] |
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| | Freund, Petra [VerfasserIn] |
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| | Meurer, Matthias [VerfasserIn] |
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| | Rahm, Johanna V. [VerfasserIn] |
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| | Knop, Michael [VerfasserIn] |
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| | Dietz, Marina S. [VerfasserIn] |
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| | Heilemann, Mike [VerfasserIn] |
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| Titel: | CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics |
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| Verf.angabe: | Tim N. Baldering, Christos Karathanasis, Marie-Lena I.E. Harwardt, Petra Freund, Matthias Meurer, Johanna V. Rahm, Michael Knop, Marina S. Dietz, and Mike Heilemann |
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| Jahr: | 2021 |
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| Jahr des Originals: | 2020 |
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| Umfang: | 20 S. |
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| Fussnoten: | Available online 7 December 2020 ; Supplemental information Seite 11-20 ; Gesehen am 07.04.2021 |
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| Titel Quelle: | Enthalten in: iScience |
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| Ort Quelle: | Amsterdam : Elsevier, 2018 |
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| Jahr Quelle: | 2021 |
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| Band/Heft Quelle: | 24(2021), 1, Artikel-ID 101895 |
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| ISSN Quelle: | 2589-0042 |
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| Abstract: | Single-molecule localization microscopy (SMLM) reports on protein organization in cells with near-molecular resolution and in combination with stoichiometric labeling enables protein counting. Fluorescent proteins allow stoichiometric labeling of cellular proteins; however, most methods either lead to overexpression or are complex and time demanding. We introduce CRISPR/Cas12a for simple and efficient tagging of endogenous proteins with a photoactivatable protein for quantitative SMLM and single-particle tracking. We constructed a HEK293T cell line with the receptor tyrosine kinase MET tagged with mEos4b and demonstrate full functionality. We determine the oligomeric state of MET with quantitative SMLM and find a reorganization from monomeric to dimeric MET upon ligand stimulation. In addition, we measured the mobility of single MET receptors in vivo in resting and ligand-treated cells. The combination of CRISPR/Cas12a-assisted endogenous protein labeling and super-resolution microscopy represents a powerful tool for cell biological research with molecular resolution. |
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| DOI: | doi:10.1016/j.isci.2020.101895 |
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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/10.1016/j.isci.2020.101895 |
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| | Volltext: https://www.sciencedirect.com/science/article/pii/S2589004220310920 |
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| | DOI: https://doi.org/10.1016/j.isci.2020.101895 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | Biochemistry |
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| | Biological Sciences Research Methodologies |
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| | Biophysics |
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| | Molecular Biology |
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| K10plus-PPN: | 1753176697 |
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| Verknüpfungen: | → Zeitschrift |
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CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics / Baldering, Tim N. [VerfasserIn]; 2021 (Online-Ressource)
