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Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.13/808663
- RNA modulators of complex phenotypes in mammalian cells
Cairns, Murray J.;
Arndt, Greg M.
- The University of Newcastle. Faculty of Health, School of Biomedical Sciences and Pharmacy
- RNA-mediated gene silencing, in the form of RNA interference (RNAi) or microRNAs (miRNAs) has provided novel tools for gene discovery and validation in mammalian cells. Here, we report on the construction and application of a random small RNA expression library for use in identifying small interfering RNA (siRNA) effectors that can modify complex cellular phenotypes in mammalian cells. The library is based in a retroviral vector and uses convergent promoters to produce unique small complementary RNAs. Using this library, we identify a range of small RNA-encoding gene inserts that overcome resistance to 5-fluorouracil (5-FU)- or tumour necrosis factor alpha (TNF-α)- induced cell death in colorectal cancer cells. We demonstrate the utility of this technology platform by identifying a key RNA effector, in the form of a siRNA, which overcomes cell death induced by the chemotherapeutic 5-FU. The technology described has the potential to identify both functional RNA modulators capable of altering physiological systems and the cellular target genes altered by these modulators.
- PLoS ONE Vol. 4, Issue 3
- Publisher Link
- Public Library of Science
- Resource Type
- journal article
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