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Science 299 (5615): 2039-2045

Copyright © 2003 by the American Association for the Advancement of Science

Identification of Hedgehog Pathway Components by RNAi in Drosophila Cultured Cells

Lawrence Lum,1 Shenqin Yao,1 Brian Mozer,2 Alessandra Rovescalli,2 Doris Von Kessler,1 Marshall Nirenberg,2 Philip A. Beachy1*

Classical genetic screens can be limited by the selectivity of mutational targeting, the complexities of anatomically based phenotypic analysis, or difficulties in subsequent gene identification. Focusing on signaling response to the secreted morphogen Hedgehog (Hh), we used RNA interference (RNAi) and a quantitative cultured cell assay to systematically screen functional roles of all kinases and phosphatases, and subsequently 43% of predicted Drosophila genes. Two gene products reported to function in Wingless (Wg) signaling were identified as Hh pathway components: a cell surface protein (Dally-like protein) required for Hh signal reception, and casein kinase 1alpha , a candidate tumor suppressor that regulates basal activities of both Hh and Wg pathways. This type of cultured cell-based functional genomics approach may be useful in the systematic analysis of other biological processes.

1 Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
2 Laboratory of Biochemical Genetics, NHLBI, NIH, Bethesda, MD 20892, USA.
*   To whom correspondence should be addressed. E-mail: pbeachy{at}jhmi.edu

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