A Combined Ex Vivo and In Vivo RNAi Screen for Notch Regulators in Drosophila Reveals an Extensive Notch Interaction Network

Abil Saj¹ - abil.saj@bsse.ethz.ch
Zeynep Arziman^3,2+ - zeynep.arziman@tubitak.gov.tr
Denise Stempfle¹⁺ - denise.stempfle@bsse.ethz.ch
Werner Van Belle¹ - werner@yellowcouch.org, werner.van.belle@gmail.com
Ursula Sauder⁴ - ursula.sauder@unibas.ch
Thomas Horn^2,3 - t.horn@dkfz.de
Markus Dürrenberger⁴ - markus.duerrenberger@unibas.ch
Renato Paro^1,5 - renato.paro@bsse.ethz.ch
Michael Boutros^3,2 - m.boutros@dkfz-heidelberg.de
Gunter Merde^1* - gunter.merdes@bsse.ethz.ch

1- Department for Biosystems Science and Engineering; ETH Zurich; Mattenstrasse 24, Building 1058, Basel; Switzerland
2- Department of Cell and Molecular Biology, University of Heidelberg, Faculty of Medicine Mannheim; 69120 Heidelberg, Germany
3- Division of Signaling and Functional Genomics, German Cancer Research Center (DKFZ);
4- Microscopy Center, Pharmazentrum, University of Basel; 4056 Basel; Switzerland
5- Faculty of Sciences; University Basel; 4056 Basel; Switzerland
* Corresponding author; + Equal Contribution

Abstract :  Notch signaling plays a fundamental role in cellular differentiation and has been linked to human diseases, including cancer. We report the use of comprehensive RNAi analyses to dissect Notch regulation and its connections to cellular pathways. A cell-based RNAi screen identified 900 candidate Notch regulators on a genome-wide scale. The subsequent use of a library of transgenic Drosophila expressing RNAi constructs enabled large-scale in vivo validation and confirmed 333 of 501 tested genes as Notch regulators. Mapping the phenotypic attributes of our data on an interaction network identified another 68 relevant genes and revealed several modules of unexpected Notch regulatory activity. In particular, we note an intriguing relationship to pyruvate metabolism, which may be relevant to cancer. Our study reveals a hitherto unappreciated diversity of tissue-specific modulators impinging on Notch and opens new avenues for studying Notch regulation and function in development and disease.

Keywords:  notch, sirna
Reference:  Abil Saj, Zeynep Arziman, Denise Stempfle, Werner Van Belle, Ursula Sauder, Thomas Horn, Markus Dürrenberger, Renato Paro, Michael Boutros, Gunter Merde; A Combined Ex Vivo and In Vivo RNAi Screen for Notch Regulators in Drosophila Reveals an Extensive Notch Interaction Network; Developmental Cell; Elsevier; volume 18; pages 862-876; May 2010
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