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This dissertation, Synthetic RNA Interference Against Influenza A Virus by Hung-chiu, Lee, 李洪釗, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled Synthetic RNA Interference Against Influenza A Virus submitted by LEE, Hung Chiu for the degree of Master of Philosophy at The University of Hong Kong In November 2005 Influenza A is one of the deadly and widespread diseases in the world. Due to its highly mutable nature of the Influenza A genome, influenza antigenicity changes rapidly, thereby making vaccination approach to have difficulties to cope with emerging viruses. In addition, vaccination fails to protect immunocompromised populations like elderly and HIV patients from infection. Anti-flu drug is therefore needed when vaccine is not suitable. Available licensed drugs either target on viral endocytosis-fusion process (e.g. M2 ion channel blocker) or virus particle released from infected cells (e.g. neuramindase inhitors). However, drug resistance against M2 ion channel blocker has been widely circulates in current H5 strains in Asia. Although neuraminidase inhibitors retain its activities against most Influenza viruses, high price and limited availability restricted its benefit to global community. As new viral protein inhibitors hunting is still expensive and tedious, different new antiviral schemes, e.g. viral messager RNA (mRNA) degradation, are actively undergone research. RNA interference (RNAi) is the one of the best studied and proven RNA degradation mechanism published nowadays. RNAi is a well-conserved mRNA degradation mechanism existed from plant to human. Viral mRNA could be destroyed by RNAi and thus suppresses virus replication. Synthetic small interfering RNA molecules (siRNA) could induce sequence specific mRNA degradation and hence efficiently inhibit target protein production. Results: This study showed that synthetic siRNA against the Influenza A virus matrix protein mRNA could reduce fusion matrix protein in human embryonic kidney cell line. Most importantly, Influenza A virus replication in MDCK cell culture system was suppressed by synthetic siRNA. Although RNAi against Influenza is subject to genome mutation, new siRNA could be redesigned to eliminate the virus quickly. Plant produced siRNA against Influenza A Non-structural Protein (NS1) is also capable to decrease virus replication. Genetically modified plant may be used as a cheap siRNA source against influenza production. Conclusions: Effective siRNA against Influenza A infection requires systemic siRNA design, careful choice of target and efficient siRNA delivery method. DOI: 10.5353/th_b3553781 Subjects: Small interfering RNAInfluenza A virusGene Silencing

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