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
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author DasGupta, Ramanuj
Nybakken, Kent
Booker, Matthew
Mathey-Prevot, Bernard
Gonsalves, Foster
Changkakoty, Binita
Perrimon, Norbert
author_facet DasGupta, Ramanuj
Nybakken, Kent
Booker, Matthew
Mathey-Prevot, Bernard
Gonsalves, Foster
Changkakoty, Binita
Perrimon, Norbert
author_sort DasGupta, Ramanuj
collection PubMed
description 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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spelling pubmed-23750412008-05-10 A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila DasGupta, Ramanuj Nybakken, Kent Booker, Matthew Mathey-Prevot, Bernard Gonsalves, Foster Changkakoty, Binita Perrimon, Norbert Genome Biol Method 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. BioMed Central 2007 2007-09-28 /pmc/articles/PMC2375041/ /pubmed/17903264 http://dx.doi.org/10.1186/gb-2007-8-9-r203 Text en Copyright © 2007 DasGupta et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Method
DasGupta, Ramanuj
Nybakken, Kent
Booker, Matthew
Mathey-Prevot, Bernard
Gonsalves, Foster
Changkakoty, Binita
Perrimon, Norbert
A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title_full A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title_fullStr A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title_full_unstemmed A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title_short A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila
title_sort case study of the reproducibility of transcriptional reporter cell-based rnai screens in drosophila
topic Method
url 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
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