| |  Online-Ressource |
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| Verfasst von: | Michels, Birgitta E. [VerfasserIn]  |
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| | Mosa, Mohammed H. [VerfasserIn] |
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| | Streibl, Barbara I. [VerfasserIn] |
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| | Zhan, Tianzuo [VerfasserIn] |
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| | Menche, Constantin [VerfasserIn] |
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| | Abou-El-Ardat, Khalil [VerfasserIn] |
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| | Darvishi, Tahmineh [VerfasserIn] |
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| | Członka, Ewelina [VerfasserIn] |
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| | Wagner, Sebastian [VerfasserIn] |
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| | Winter, Jan [VerfasserIn] |
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| | Medyouf, Hind [VerfasserIn] |
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| | Boutros, Michael [VerfasserIn] |
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| | Farin, Henner F. [VerfasserIn] |
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| Titel: | Pooled in vitro and in vivo CRISPR-Cas9 screening identifies tumor suppressors in human colon organoids |
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| Verf.angabe: | Birgitta E. Michels, Mohammed H. Mosa, Barbara I. Streibl, Tianzuo Zhan, Constantin Menche, Khalil Abou-El-Ardat, Tahmineh Darvishi, Ewelina Członka, Sebastian Wagner, Jan Winter, Hind Medyouf, Michael Boutros, Henner F. Farin |
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| E-Jahr: | 2020 |
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| Jahr: | 7 May 2020 |
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| Umfang: | 11 S. |
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| Fussnoten: | Gesehen am 10.06.2020 |
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| Titel Quelle: | Enthalten in: Cell stem cell |
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| Ort Quelle: | Amsterdam [u.a.] : Elsevier, 2007 |
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| Jahr Quelle: | 2020 |
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| Band/Heft Quelle: | 26(2020), 5, Seite 782-792 |
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| ISSN Quelle: | 1875-9777 |
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| Abstract: | Colorectal cancer (CRC) is characterized by prominent genetic and phenotypic heterogeneity between patients. To facilitate high-throughput genetic testing and functional identification of tumor drivers, we developed a platform for pooled CRISPR-Cas9 screening in human colon organoids. Using transforming growth factor β (TGF-β) resistance as a paradigm to establish sensitivity and scalability in vitro, we identified optimal conditions and strict guide RNA (gRNA) requirements for screening in 3D organoids. We then screened a pan-cancer tumor suppressor gene (TSG) library in pre-malignant organoids with APC−/−;KRASG12D mutations, which were xenografted to study clonal advantages in context of a complex tumor microenvironment. We identified TGFBR2 as the most prevalent TSG, followed by known and previously uncharacterized mediators of CRC growth. gRNAs were validated in a secondary screen using unique molecular identifiers (UMIs) to adjust for clonal drift and to distinguish clone size and abundance. Together, these findings highlight a powerful organoid-based platform for pooled CRISPR-Cas9 screening for patient-specific functional genomics. |
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| DOI: | doi:10.1016/j.stem.2020.04.003 |
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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.stem.2020.04.003 |
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| | Volltext: http://www.sciencedirect.com/science/article/pii/S1934590920301429 |
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| | DOI: https://doi.org/10.1016/j.stem.2020.04.003 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | clonal drift |
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| | colorectal cancer |
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| | human colonic stem cells |
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| | lentiviral barcoding |
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| | non-homologous end joining |
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| | patient-derived organoids |
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| | pooled-barcoded CRISPR-Cas9 screening |
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| | tumor microenvironment |
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| | tumor suppressor genes |
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| | unique molecular identifiers |
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| K10plus-PPN: | 1700352989 |
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| Verknüpfungen: | → Zeitschrift |
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Pooled in vitro and in vivo CRISPR-Cas9 screening identifies tumor suppressors in human colon organoids / Michels, Birgitta E. [VerfasserIn]; 7 May 2020 (Online-Ressource)
