Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families
Signaling pathway modules are often encoded by several closely related paralogous genes that can have redundant roles and are therefore difficult to analyze by loss-of-function analysis. A typical example is the Wnt signaling pathway, which in mammals is mediated by 19 Wnt ligands that can bind to 1...
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| Hauptverfasser: | , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
21 Jul 2017
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| In: |
The FASEB journal
Year: 2017, Jahrgang: 31, Heft: 11, Pages: 4832-4844 |
| ISSN: | 1530-6860 |
| DOI: | 10.1096/fj.201700144R |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1096/fj.201700144R Verlag, Volltext: https://www-fasebj-org.ezproxy.medma.uni-heidelberg.de/doi/10.1096/fj.201700144R 
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| Verfasserangaben: | Oksana Voloshanenko, Philipp Gmach, Jan Winter, Dominique Kranz, and Michael Boutros |
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| 245 | 1 | 0 | |a Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families |c Oksana Voloshanenko, Philipp Gmach, Jan Winter, Dominique Kranz, and Michael Boutros |
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| 520 | |a Signaling pathway modules are often encoded by several closely related paralogous genes that can have redundant roles and are therefore difficult to analyze by loss-of-function analysis. A typical example is the Wnt signaling pathway, which in mammals is mediated by 19 Wnt ligands that can bind to 10 Frizzled (FZD) receptors. Although significant progress in understanding Wnt-FZD receptor interactions has been made in recent years, tools to generate systematic interaction maps have been largely lacking. Here we generated cell lines with multiplex mutant alleles of FZD1, FZD2, and FZD7 and demonstrate that these cells are unresponsive to canonical Wnt ligands. Subsequently, we performed genetic rescue experiments with combinations of FZDs and canonical Wnts to create a functional ligand-receptor interaction map. These experiments showed that whereas several Wnt ligands, such as Wnt3a, induce signaling through a broad spectrum of FZD receptors, others, such as Wnt8a, act through a restricted set of FZD genes. Together, our results map functional interactions of FZDs and 10 Wnt ligands and demonstrate how multiplex targeting by clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 can be used to systematically elucidate the functions of multigene families.—Voloshanenko, O., Gmach, P., Winter, J., Kranz, D., Boutros, M. Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families. | ||
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