A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila.
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.
DasGupta
Ramanuj
R
New York University School of Medicine/Cancer Institute, Department of Pharmacology, First Avenue, New York, NY 10016, USA. ramanuj.dasgupta@med.nyu.edu
Nybakken
Kent
K
Booker
Matthew
M
Mathey-Prevot
Bernard
B
Gonsalves
Foster
F
Changkakoty
Binita
B
Perrimon
Norbert
N
D000818:Animals; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D015723:Gene Library; D017344:Genes, Insect; D005821:Genetic Techniques; D016678:Genome; D008957:Models, Genetic; D012313:RNA / Q000235:genetics; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D029721:Drosophila Proteins; D012330:RNA, Double-Stranded; D012313:RNA