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Applications of RNA interference high-throughput screening technology in cancer biology and virology.

Gao S, Yang C, Jiang S, Xu XN, Lu X, He YW, Cheung A, Wang H - Protein Cell (2014)

Bottom Line: RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function.This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight.Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, John Radcliffe Hospital, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK, shan.gao@oncology.ox.ac.uk.

ABSTRACT
RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function. Large-scale gene silencing using RNAi high-throughput screening (HTS) has opened an exciting frontier to systematically study gene function in mammalian cells. This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight. Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.

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Related in: MedlinePlus

Work flow of genome wide siRNA screen for viral related host factor. Cells are initially transfected with different siRNA libraries, and then infected with viruses. Different assays can be utilized to detect screen results, including luciferase reporter assay, Tat-dependent reporter assay, p24 detection, as well as GFP detection. After first round of screen, hit analysis is performed and the candidates are subject to additional screen with individual siRNAs
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Fig3: Work flow of genome wide siRNA screen for viral related host factor. Cells are initially transfected with different siRNA libraries, and then infected with viruses. Different assays can be utilized to detect screen results, including luciferase reporter assay, Tat-dependent reporter assay, p24 detection, as well as GFP detection. After first round of screen, hit analysis is performed and the candidates are subject to additional screen with individual siRNAs

Mentions: The Drosophila C virus (DCV), a picornavirus, was the first virus to undergo genome-wide RNAi screening to identify novel host factors required for internal ribosome entry site (IRES)-dependent viral translation (Cherry et al., 2005). The first virus in a mammalian system that was screened by genome wide RNAi is HCV in 2007. In an effort to discover novel druggable targets against HCV, a library of siRNAs targeting 4,000 human genes was used to identify genes that regulate HCV replication in Huh7-derived EN5-3 cells harboring an HCV subgenomic replicon (Fig. 3). Nine cellular genes, including members of the tumor necrosis factor/lymphotoxin signaling pathway, were identified (Table 1). Silencing these genes leads to inhibition of HCV replication, and the level of siRNA silencing of these host genes correlates well with the inhibition of HCV (Ng et al., 2007).Figure 3


Applications of RNA interference high-throughput screening technology in cancer biology and virology.

Gao S, Yang C, Jiang S, Xu XN, Lu X, He YW, Cheung A, Wang H - Protein Cell (2014)

Work flow of genome wide siRNA screen for viral related host factor. Cells are initially transfected with different siRNA libraries, and then infected with viruses. Different assays can be utilized to detect screen results, including luciferase reporter assay, Tat-dependent reporter assay, p24 detection, as well as GFP detection. After first round of screen, hit analysis is performed and the candidates are subject to additional screen with individual siRNAs
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4225462&req=5

Fig3: Work flow of genome wide siRNA screen for viral related host factor. Cells are initially transfected with different siRNA libraries, and then infected with viruses. Different assays can be utilized to detect screen results, including luciferase reporter assay, Tat-dependent reporter assay, p24 detection, as well as GFP detection. After first round of screen, hit analysis is performed and the candidates are subject to additional screen with individual siRNAs
Mentions: The Drosophila C virus (DCV), a picornavirus, was the first virus to undergo genome-wide RNAi screening to identify novel host factors required for internal ribosome entry site (IRES)-dependent viral translation (Cherry et al., 2005). The first virus in a mammalian system that was screened by genome wide RNAi is HCV in 2007. In an effort to discover novel druggable targets against HCV, a library of siRNAs targeting 4,000 human genes was used to identify genes that regulate HCV replication in Huh7-derived EN5-3 cells harboring an HCV subgenomic replicon (Fig. 3). Nine cellular genes, including members of the tumor necrosis factor/lymphotoxin signaling pathway, were identified (Table 1). Silencing these genes leads to inhibition of HCV replication, and the level of siRNA silencing of these host genes correlates well with the inhibition of HCV (Ng et al., 2007).Figure 3

Bottom Line: RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function.This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight.Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, John Radcliffe Hospital, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK, shan.gao@oncology.ox.ac.uk.

ABSTRACT
RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function. Large-scale gene silencing using RNAi high-throughput screening (HTS) has opened an exciting frontier to systematically study gene function in mammalian cells. This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight. Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.

Show MeSH
Related in: MedlinePlus