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Multiplexing siRNAs to compress RNAi-based screen size in human cells

  1. Author:
    Martin, S. E.
    Jones, T. L.
    Thomas, C. L.
    Lorenzi, P. L.
    Nguyen, D. A.
    Runfola, T.
    Gunsior, M.
    Weinstein, J. N.
    Goldsmith, P. K.
    Lader, E.
    Huppi, K.
    Caplen, N. J.

  2. Author Address

    NCI, Gene Silencing Sect, Off Sci & Technol Partnership, OD,CCR,NIH, Bethesda, MD 20892 USA. NCI Frederick, Mol Target Dev Program, CCR, NIH, Frederick, MD USA. NCI, Genom & Bioinformat Grp, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA. NCI, CCR, Antibody & Prot Purif Unit, NIH, Bethesda, MD 20892 USA. Qiagen Inc, Germantown, MD USA.;Caplen, NJ, NCI, Gene Silencing Sect, Off Sci & Technol Partnership, OD,CCR,NIH, Bethesda, MD 20892 USA.;ncaplen@mail.nih.gov
    1. Year: 2007
    2. Date: Apr
  2. Journal: Nucleic Acids Research
    1. 35
    2. 8
  4. Type of Article: Article
  5. Article Number: e57
  6. ISSN: 0305-1048
  1. Abstract:

    Here we describe a novel strategy using multiplexes of synthetic small interfering RNAs (siRNAs) corresponding to multiple gene targets in order to compress RNA interference (RNAi) screen size. Before investigating the practical use of this strategy, we first characterized the gene-specific RNAi induced by a large subset (258 siRNAs, 129 genes) of the entire siRNA library used in this study (similar to 800 siRNAs, similar to 400 genes). We next demonstrated that multiplexed siRNAs could silence at least six genes to the same degree as when the genes were targeted individually. The entire library was then used in a screen in which randomly multiplexed siRNAs were assayed for their affect on cell viability. Using this strategy, several gene targets that influenced the viability of a breast cancer cell line were identified. This study suggests that the screening of randomly multiplexed siRNAs may provide an important avenue towards the identification of candidate gene targets for downstream functional analyses and may also be useful for the rapid identification of positive controls for use in novel assay systems. This approach is likely to be especially applicable where assay costs or platform limitations are prohibitive.

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  2. DOI: 10.1093/nar/gkm141
  3. View record in Web of Science®
  4. WOS: 000247239600035

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