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A Case Study of the Reproducibility of Transcriptional Reporter Cell-Based RNAi Screens in Drosophila

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2007

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BioMed Central
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DasGupta, Ramanuj, Kent Nybakken, Matthew Booker, Bernard Mathey-Prevot, Foster Gonsalves, Binita Changkakoty, and Norbert Perrimon. 2007. A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila. Genome Biology 8(9): R203.

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Abstract

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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