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 s...

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Detalles Bibliográficos
Autores principales: DasGupta, Ramanuj, Nybakken, Kent, Booker, Matthew, Mathey-Prevot, Bernard, Gonsalves, Foster, Changkakoty, Binita, Perrimon, Norbert
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2007
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2375041/
https://www.ncbi.nlm.nih.gov/pubmed/17903264
http://dx.doi.org/10.1186/gb-2007-8-9-r203
Descripción
Sumario: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.

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