Project Z03

Project leader: Michael Boutros

Advanced genetic screens and genome engineering

The Z03 Central Project provides access to screening approaches including CRISPR-Cas9 and RNAi screening in cell culture models, as well as time- and spatially-resolved CRISPR and RNAi screening in Drosophila. The Z03 project generates cell lines in which Wnt signaling pathway components are endogenously modified by fluorescent proteins using genome editing to perform quantitative analysis using advanced imaging techniques. The platform supports screening procedures in cell culture models and generates a collection of Drosophila CRISPR lines for tissue-specific in vivo studies.

Hochentwickelte genetische Screens und Genom-Editierung

Das Z03-Zentralprojekt bietet Zugang zu Screening-Ansätzen einschließlich CRISPR-Cas9 und RNAi-Screening in Zellkulturmodellen, sowie zeit- und räumlich aufgelöstes CRISPR und RNAi-Screening in Drosophila. Im Rahmen des Z03-Projektes werden Zelllinien generiert, in dem Wnt-Signalwegskomponenten endogen durch Fluoreszenzproteine mittels Genom-Editierung modifiziert werden, um quantitative Untersuchungen mittels Mikroskopie-Verfahren durchzuführen. Die Plattform unterstützt Screening-Verfahren in Zellkulturmodellen und generiert eine Kollektion von Drosophila CRISPR-Linien für gewebespezifische in vivo Studien.

Project-related Publications

  • Ambrosi G., O. Voloshanenko, A.F. Eckert, D. Kranz, G.U. Nienhaus, and M. Boutros. 2020. Allele-specific endogenous tagging and quantitative analysis of β-catenin in colon cancer cells. BioRxiv, https://doi.org/10.1101/2020.06.18.159616
  • Imkeller, K., G. Ambrosi, M. Boutros, and W. Huber. 2020. gscreend: modelling asymmetric count ratios in CRISPR screens to decrease experiment size and improve phenotype detection. Genome Biol. 21:53. PMID: 32122365
  • Lin, Y.C., A. Haas, A. Bufe, S. Parbin, M. Hennecke, O. Voloshanenko, J.C. Gross, M. Boutros, S.P. Acebron, and H. Bastians. 2020. Wnt10b-GSK3β-dependent Wnt/STOP signaling prevents aneuploidy in human somatic cells. Life Science Alliance. 4(1):e202000855. PMID: 33257473
  • Port, F., M. Starostecka, and M. Boutros. 2020. Multiplexed conditional genome editing with Cas12a in Drosophila. PNAS 22890:22899. PMID: 32843348
  • Port, F., C. Strein, M. Stricker, B. Rauscher, F. Heigwer, J. Zhou, C. Beyersdorffer, J. Frei, A. Hess, K. Kern, L. Lange, N. Langner, R. Malamud, B. Pavlovic, K. Radecke, L. Schmitt, L. Voos, E. Valentini, and M. Boutros. 2020. A large-scale resource for tissue-specific CRISPR mutagenesis in Drosophila. eLife. 9. PMID: 32053108
  • Wesslowski, J., P. Kozielewicz, X. Wang, H. Cui, H. Schihada, D. Kranz, M.P. Karuna, P. Levkin, J.C. Gross, M. Boutros, G. Schulte, and G. Davidson. 2020. eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled. J Biol Chem. 295:8759-8774. PMID: 32381507
  • Chaudhary C., S. Hingole, J. Frei, F. Port, D. Strutt, and M. Boutros. 2019. Robust Wnt signaling is maintained by a Wg protein gradient and Fz2 receptor activity in the developing Drosophila wing. Development. 46(15):dev174789.
  • Rauscher B., F. Heigwer, L. Henkel, T. Hielscher, and M. Boutros. 2018. Toward an integrated map of genetic interactions in cancer cells. Mol Syst Biol. e7656. PMID: 29467179
  • Voloshanenko, O., U. Schwartz, D. Kranz, B. Rauscher, M. Linnebacher, I. Augustin, and M. Boutros. 2018. beta-catenin-independent regulation of Wnt target genes by RoR2 and ATF2/ATF4 in colon cancer cells. Sci Rep. 8:3178. PMID: 29453334
  • Voloshanenko, O., P. Gmach, J. Winter, D. Kranz, and M. Boutros. 2017. Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families. FASEB J. 31:4832-4844. PMID: 28733458