|
|
Multiplexing siRNAs to compress RNAi-based screen size in human cells
Martin SE, Jones TL, Thomas CL, Lorenzi PL, Nguyen DA, Runfola T, Gunsior M, Weinstein JN,
Goldsmith PK, Lader E, Huppi K, Caplen NJ
Nucleic Acids Res. 2007 Mar 28
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 (~800 siRNAs, ~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.
|
