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Proc Natl Acad Sci U S A. 2017 Dec 12;114(50):E10755-E10762. doi: 10.1073/pnas.1713362114. Epub 2017 Nov 28.

Improved detection of synthetic lethal interactions in Drosophila cells using variable dose analysis (VDA).

Author information

1
Living Systems Institute, University of Exeter Medical School, University of Exeter, Exeter, United Kingdom EX4 4QD; b.housden@exeter.ac.uk perrimon@genetics.med.harvard.edu.
2
Department of Genetics, Harvard Medical School, Boston, MA 02115.
3
Department of Genetics and Complex Diseases, Harvard T. H. Chan School of Public Health, Boston, MA 02115.
4
Department of Genetics, Harvard Medical School, Boston, MA 02115; b.housden@exeter.ac.uk perrimon@genetics.med.harvard.edu.
5
Howard Hughes Medical Institute, Boston, MA 02115.

Abstract

Synthetic sick or synthetic lethal (SS/L) screens are a powerful way to identify candidate drug targets to specifically kill tumor cells, but this approach generally suffers from low consistency between screens. We found that many SS/L interactions involve essential genes and are therefore detectable within a limited range of knockdown efficiency. Such interactions are often missed by overly efficient RNAi reagents. We therefore developed an assay that measures viability over a range of knockdown efficiency within a cell population. This method, called Variable Dose Analysis (VDA), is highly sensitive to viability phenotypes and reproducibly detects SS/L interactions. We applied the VDA method to search for SS/L interactions with TSC1 and TSC2, the two tumor suppressors underlying tuberous sclerosis complex (TSC), and generated a SS/L network for TSC. Using this network, we identified four Food and Drug Administration-approved drugs that selectively affect viability of TSC-deficient cells, representing promising candidates for repurposing to treat TSC-related tumors.

KEYWORDS:

Drosophila; RNAi; drug target discovery; high-throughput screening; synthetic lethality

PMID:
29183982
PMCID:
PMC5740648
DOI:
10.1073/pnas.1713362114
[Indexed for MEDLINE]
Free PMC Article

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