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Genes 2012, 3(4), 686-701; doi:10.3390/genes3040686
Review

Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation

1,* , 2
, 3
 and 1,*
Received: 13 August 2012; in revised form: 27 September 2012 / Accepted: 30 October 2012 / Published: 2 November 2012
(This article belongs to the Special Issue RNA Interference)
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Abstract: RNAi is a convenient, widely used tool for screening for genes of interest. We have recently used this technology to screen roughly 750 candidate genes, in C. elegans, for potential roles in regulating muscle protein degradation in vivo. To maximize confidence and assess reproducibility, we have only used previously validated RNAi constructs and have included time courses and replicates. To maximize mechanistic understanding, we have examined multiple sub-cellular phenotypes in multiple compartments in muscle. We have also tested knockdowns of putative regulators of degradation in the context of mutations or drugs that were previously shown to inhibit protein degradation by diverse mechanisms. Here we discuss how assaying multiple phenotypes, multiplexing RNAi screens with use of mutations and drugs, and use of bioinformatics can provide more data on rates of potential false positives and negatives as well as more mechanistic insight than simple RNAi screening.
Keywords: RNAi; systems biology; network biology; functional genomics; muscle; proteolysis; C. elegans RNAi; systems biology; network biology; functional genomics; muscle; proteolysis; C. elegans
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Lehmann, S.; Shephard, F.; Jacobson, L.A.; Szewczyk, N.J. Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation. Genes 2012, 3, 686-701.

AMA Style

Lehmann S, Shephard F, Jacobson LA, Szewczyk NJ. Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation. Genes. 2012; 3(4):686-701.

Chicago/Turabian Style

Lehmann, Susann; Shephard, Freya; Jacobson, Lewis A.; Szewczyk, Nathaniel J. 2012. "Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation." Genes 3, no. 4: 686-701.



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