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mRNA Processing pp 235–245Cite as

Genome-Wide RNAi Screens for RNA Processing Events in Drosophila melanogaster S2 Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1648))

Abstract

Over the past 10 years, the design and application of genome-wide screening (GWS) has improved to the point that it can now be done at level of the individual laboratory. The advantages of GWSs compared to classical genetic screens include: immediate identification of a positive scoring gene, relatively short period of time necessary to conduct the screen (as little as 1 week), cell lines do not present developmental needs for gene expression that an organism normally would, and validation/confirmation of results is straightforward. Here, we describe a general protocol for GWS to be conducted in Drosophila melanogaster S2 cells. We provide specific details on what type of experiments must be done before initiating a screen, the materials that are required to conduct a screen, and make suggestions on methods to carry out secondary screening and counter-screening once the initial GWS is complete. Multiple considerations are also raised that focus on how to anticipate false positives/negatives and how to minimize their occurrence through intelligent design. Finally, we provide specific examples of data that our group has gathered from published genome-wide screens in order to exemplify how “hits” are scored and confirmed.

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Author information

Authors and Affiliations

  1. Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Natoya Peart

  2. Department of Biochemistry and Molecular Biology, University of Texas Medical Branch at Galveston, 301 University Blvd., Galveston, TX, 77550, USA

    Eric J. Wagner

Authors
  1. Natoya Peart
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  2. Eric J. Wagner
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Corresponding author

Correspondence to Eric J. Wagner .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, California, USA

    Yongsheng Shi

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Peart, N., Wagner, E.J. (2017). Genome-Wide RNAi Screens for RNA Processing Events in Drosophila melanogaster S2 Cells. In: Shi, Y. (eds) mRNA Processing. Methods in Molecular Biology, vol 1648. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7204-3_17

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  • DOI: https://doi.org/10.1007/978-1-4939-7204-3_17

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7203-6

  • Online ISBN: 978-1-4939-7204-3

  • eBook Packages: Springer Protocols

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