Key Points
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RNA interference (RNAi) allows for the specific silencing of genes. With the development of RNAi libraries, the effects of inhibiting specific genes can now be carried out in a high-throughput and systematic manner.
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We survey the pros and cons of existing RNAi libraries and screening strategies, and also suggest the salient points to be considered when designing an RNAi screen.
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RNAi screening is now showing potential as an invaluable tool in the process of target identification.
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RNAi is also showing promise as a useful tool in target validation.
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Although RNAi has already been used in target identification and validation processes, we suggest that it can also be used in the stages of compound identification and lead optimization.
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There is particular utility in using RNAi to generate gene-expression signatures that characterize the specific inhibition of a potential drug target. These signatures can be compared to gene expression signatures from small-molecule inhibitors and can be used to identify more specific compounds. The signatures can also be used to understand the mechanism of action of small-molecule inhibitors.
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Although RNAi is not a panacea to all of the rate-limiting steps of drug discovery, it does have potential in certain areas of this process. We discuss the limitations of RNAi screening and in particular, the problem of off-target effects.
Abstract
With the development of RNA interference (RNAi) libraries, systematic and cost-effective genome-wide loss-of-function screens can now be carried out with the aim of assessing the role of specific genes in neoplastic phenotypes, and the rapid identification of novel drug targets. Here, we discuss the existing applications of RNAi in cancer drug discovery and highlight areas in this process that may benefit from this technology in the future.
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Glossary
- Glioblastoma
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A high-grade brain malignancy arising from astrocytes with abnormal cellular proliferation and increased tumour angiogenesis. This cancer is usually refractory to chemotherapy and has a poor prognosis.
- Isogenic-paired cell lines
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These consist of two cell lines that are of identical origin, but differ at a known genetic locus.
- Chemosensitization
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The ability of a defined change to increase the sensitivity of a cell to a chemotherapeutic agent.
- CRE-loxP system
-
Murine strains can be engineered with loxP sites flanking a gene of interest. Expression of CRE recombinase, an enzyme that causes selective excision of all genetic material between two loxP sites, allows for ablation of the gene of interest. Expression of CRE under the control of different promoter elements enables temporal and tissue-specific deletion of the gene of interest.
- Tet-off system
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Murine strains can be engineered to contain a transgene of interest that is either induced (tet-ON) or repressed (tet-OFF) by tetracycline analogues (such as doxycycline). An advantage of tet-regulation is that the gene of interest can be serially induced or repressed by withdrawing or adding doxycycline to the animal's drinking water.
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Iorns, E., Lord, C., Turner, N. et al. Utilizing RNA interference to enhance cancer drug discovery. Nat Rev Drug Discov 6, 556–568 (2007). https://doi.org/10.1038/nrd2355
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DOI: https://doi.org/10.1038/nrd2355
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