title: Reagent and Data Resources for Investigation of RNA Binding Protein Functions in Drosophila melanogaster Cultured Cells.
abstract: RNA binding proteins (RBPs) are involved in many cellular functions. To facilitate functional characterization of RBPs, we generated an RNA interference (RNAi) library for Drosophila cell-based screens comprising reagents targeting known or putative RBPs. To test the quality of the library and provide a baseline analysis of the effects of the RNAi reagents on viability, we screened the library using a total ATP assay and high-throughput imaging in Drosophila S2R+ cultured cells. The results are consistent with production of a high-quality library that will be useful for functional genomics studies using other assays. Altogether, we provide resources in the form of an initial curated list of Drosophila RBPs; an RNAi screening library we expect to be used with additional assays that address more specific biological questions; and total ATP and image data useful for comparison of those additional assay results with fundamental information such as effects of a given reagent in the library on cell viability. Importantly, we make the baseline data, including more than 200,000 images, easily accessible online.
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115 stephanie_mohr@hms.harvard.edu.
authors.forename: Stephanie E
authors.initials: SE
authors.lastname: Mohr
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Kirstin
authors.initials: K
authors.lastname: Rudd
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Michael
authors.initials: M
authors.lastname: Buckner
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Quentin
authors.initials: Q
authors.lastname: Gilly
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Blake
authors.initials: B
authors.lastname: Foster
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Sierzputowska
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Aram
authors.initials: A
authors.lastname: Comjean
authors.affiliation: Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Bing
authors.initials: B
authors.lastname: Ye
authors.affiliation: Drosophila RNAi Screening Center, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115 Howard Hughes Medical Institute, Boston, Massachusetts 02115.
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D019295:Computational Biology / Q000379:methods; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D023281:Genomics / Q000379:methods; D057166:High-Throughput Screening Assays; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D000255:Adenosine Triphosphate
keywords: Drosophila; RNA binding protein; RNAi; data sharing; high-throughput screen