A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila

Genome Biol. 2007;8(9):R203. doi: 10.1186/gb-2007-8-9-r203.


Off-target effects have been demonstrated to be a major source of false-positives in RNA interference (RNAi) high-throughput screens. In this study, we re-assess the previously published transcriptional reporter-based whole-genome RNAi screens for the Wingless and Hedgehog signaling pathways using second generation double-stranded RNA libraries. Furthermore, we investigate other factors that may influence the outcome of such screens, including cell-type specificity, robustness of reporters, and assay normalization, which determine the efficacy of RNAi-knockdown of target genes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Drosophila / genetics*
  • Drosophila Proteins / metabolism
  • Gene Expression Regulation
  • Gene Library
  • Genes, Insect
  • Genetic Techniques
  • Genome
  • Models, Genetic
  • RNA / genetics
  • RNA Interference*
  • RNA, Double-Stranded / genetics
  • Sensitivity and Specificity
  • Transcription, Genetic


  • Drosophila Proteins
  • RNA, Double-Stranded
  • RNA