using multiple phenotype assays and epistasis testing to enhance the reliability of rnai screening and identify regulators of muscle protein degradation使用多个表型分析和上位测试提高rnai的可靠性筛选和识别监管者肌肉蛋白质的降解.pdfVIP

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using multiple phenotype assays and epistasis testing to enhance the reliability of rnai screening and identify regulators of muscle protein degradation使用多个表型分析和上位测试提高rnai的可靠性筛选和识别监管者肌肉蛋白质的降解.pdf

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using multiple phenotype assays and epistasis testing to enhance the reliability of rnai screening and identify regulators of muscle protein degradation使用多个表型分析和上位测试提高rnai的可靠性筛选和识别监管者肌肉蛋白质的降解

Genes 2012, 3, 686-701; doi:10.3390/genes3040686 OPEN ACCESS genes ISSN 2073-4425 /journal/genes Review Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation Susann Lehmann 1,*, Freya Shephard 2, Lewis A. Jacobson 3 and Nathaniel J. Szewczyk 1,* 1 Medical Resarch Council/Arthritis Research UK Centre for Musculoskeletal Ageing Research, University of Nottingham, Royal Derby Hospital, Derby, DE22 3DT, UK 2 Medical Resarch Council/Arthritis Research UK Centre for Musculoskeletal Ageing Research, University of Nottingham, School of Veterinary Medicine Science, Sutton Bonington, LE12 5RD, UK; E-Mail: Freya.Shephard@nottingham.ac.uk 3 Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA; E-Mail: ljac@ * Authors to whom correspondence should be addressed; E-Mails: lehmann.susann@ (S.L.); nate@ (N.S.); Tel.: +44-1332-724615; Fax: +44-1332-724727. Received: 13 August 2012; in revised form: 27 September 2012 / Accepted: 30 October 2012 / Published: 2 November 2012 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.