Developing Anti-viral Ribozymes to Suppress Arboviruses in Transgenic Mosquitoes

描述（由申请人提供）：基孔肯雅热（CHIKV）、黄热病（YFV）和登革热（DENV）是最棘手的人类健康病原体之一，每年造成数亿人感染和数十万人死亡。它们通过单一蚊子种类埃及伊蚊传播给人类。有效的人用疫苗尚不适用于CHIK和DENV，尽管有有效的疫苗，但YFV作为致命病原体的重新出现表明持续根除的疫苗接种策略无效。干扰节肢动物媒介中的虫媒病毒孵化作为中断病毒传播周期和降低向人类传播效率的一种方式正在受到相当大的关注。其中一种方法设想用对感染有抵抗力的蚊子取代易感蚊子，或阻止疾病传播。这种方法具有环境安全、成本效益和长期抑制疾病的明显优势。转基因引入表达的抗病毒分子以在蚊子载体中产生抗性现在是真实的可能性。最近的成功反映在RIDL的方法显示出显着的承诺，可以合理地预期再加上转基因的耐火性，建立和维持转基因效应在反弹蚊子种群。然而，这一途径必须与确保长期有效性和稳定的转基因表达相结合。我们的研究表明，核酶（RNA-酶）介导的病毒抑制可以提供一种有效的抗病毒转基因免疫手段。最近的努力已经证实了锤头状核酶和I组内含子在抑制蚊子细胞中的DENV感染中的有效性。这项研究的目的是建立在我们成功开发抗病毒核酶作为有效的效应基因对虫媒病毒感染的蚊子。特别是，该提案将专注于开发，优化和验证有效的核酶抑制策略，以对抗由相同蚊子物种，埃及伊蚊，DENV，CHIKV和YF传播的三种主要新兴和重新出现的疾病病原体。最终目标是能够提供可用于同时消除两种或更多种这些病毒性疾病的蚊子菌株。 公共卫生相关性：基孔肯雅热（CHIKV）、黄热病（YFV）和登革热（DENV）是最麻烦的单蚊传播的人类健康病原体之一，每年造成数亿人感染和数十万人死亡。这项拟议的研究旨在开发抗病毒核酶，作为抑制这些虫媒病毒在蚊子埃及伊蚊中感染的手段，从而防止疾病传播。

DESCRIPTION (provided by applicant): Chikungunya (CHIKV), yellow fever (YFV), and Dengue (DENV) are among the most troublesome human health pathogens responsible for hundreds of millions of infections and hundreds of thousands of deaths each year. They are transmitted to human populations by a single mosquito species, Aedes aegypti. Effective human vaccines are not yet available for CHIK, and DENV, and the re-emergence of YFV as a lethal pathogen, in spite of the availability of an effective vaccine, suggests ineffectiveness of vaccination strategies for sustained eradication. Interference with the incubation arboviruses within the arthropod vector is receiving considerable attention as a way to interrupt the viral transmission cycle and reduce the efficiency of transmission to humans. One such approach envisions population replacement of susceptible mosquitoes with those resistant to infection or to halt disease transmission. This approach has distinct advantages of environmentally safety, cost effectiveness, and long-term disease suppression. Transgenic introduction of expressed anti-viral molecules to generate refractoriness in the mosquito vector is now a real possibility. Recent successes reflected in the RIDL approach show significant promise, and can reasonably be expected to be coupled with transgenic refractoriness to establish and maintain transgene effectors in rebound mosquito populations. However, this avenue must necessarily be coupled with ensuring both long term effectiveness and stable transgene expression. Our research is demonstrating that ribozyme (RNA-enzyme) mediated viral suppression can provide an effective means of transgenic immunization against viruses. Recent efforts have confirmed the effectiveness of hammerhead ribozymes and Group I introns in suppressing DENV infection in mosquito cells. The research proposed seeks to build upon our successes in developing anti-viral ribozymes as potent effector genes against arbovirus infections in mosquitoes. In particular, this proposal will focus on developing, optimizing, and validating effective ribozyme suppression strategies against three major emerging and re-emerging disease pathogens vectored by the same mosquito species, Aedes aegypti, DENV, CHIKV, and YF. The ultimate goal is to be able to provide mosquito strains that will be useful in simultaneously eliminating two or more of these viral diseases. PUBLIC HEALTH RELEVANCE: Chikungunya (CHIKV), yellow fever (YFV), and Dengue (DENV) are among the most troublesome single-mosquito transmitted human health pathogens causing hundreds of millions of infections and hundreds of thousands of deaths each year. This proposed research seeks to develop anti-viral ribozymes as means of suppressing the infection of these arbovirus in the mosquito, Aedes aegypti, thereby preventing disease transmission.