New Alphavirus Expression Systems in Mosquitoes

蚊子中的新甲病毒表达系统

基本信息

批准号：
8312695
负责人：
KENNETH E OLSON
金额：
$ 28.22万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-04-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8312695
关键词：
Adult Aedes Affect Alphavirus Alphavirus Infections Animal Model Animals Anopheles Genus Anopheles gambiae Anti-Bacterial Agents Antibodies Antiviral Agents Antiviral Response Applications Grants Arboviruses Bioinformatics Biological Markers Biology Bioluminescence Cells Cessation of life Competence Complex Coupled Culex (Genus)Culex pipiens Culicidae Detection Development Double-Stranded RNA Engineering Evaluation Explosion Fat Body Fluorescence Gene Expression Gene Targeting Gene Transfer Techniques Genes Genetic Genetic Transcription Genome Genomics Goals Grant Health Hemocytes Human Imagery Immune Infection Injection of therapeutic agent Insecta Institution Lead Life Luciferases Methods Midgut Modeling Molecular Morbidity - disease rate Mosquito-borne infectious disease Mus Neurotoxins Pathogenesis Pathway interactions Peptides Pharmaceutical Preparations Phase Polymerase Positioning Attribute Proteins Protocols documentation RNA RNA Interference RNA Interference Pathway RNA Virus Infections RNA Viruses Reporter Reporter Genes Research Research Infrastructure Research Personnel Resistance Route Salivary Glands Small RNA Surface System Techniques Technology Testing Time Togaviridae Transgenes Transgenic Organisms Vaccines Virion Virus Virus Diseases Virus Replication Western Equine Encephalitis Virus animal care base chikungunya design disease transmission genome sequencing improved interest knock-down luciferin mortality mouse model novel novel vaccines pathogen site-specific integration tool transmission process vector vector transmission

项目摘要

DESCRIPTION (provided by applicant): Mosquito-borne viral diseases remain significant causes of morbidity and mortality throughout much of the world and impose tremendous burdens on human and animal care infrastructures. It is critical that we have a better understanding of the molecular basis of vector competence and arbovirus transmission by mosquitoes. Genome sequence information and other bioinformatics are available for three major vector species (Anopheles gambiae, Aedes aegypti and Culex pipiens). New techniques are still needed by vector biologists and arbovirologists to fully exploit this explosion of information and develop a more complete understanding of mosquito-borne disease transmission. For the last 15 years, AIDL researchers have been at the forefront developing molecular tools termed alphavirus transducing systems or ATS's that allow gene expression in a variety of mosquito (and insect) species. ATS's have facilitated gene expression in adult mosquitoes, allowed functional analyses of vector and alphavirus genes, and have been used in the study of mosquito-alphavirus interactions. Vector biologists at AIDL, and elsewhere, are currently manipulating innate immune pathways in mosquitoes and need tools such as ATS's to characterize the effects of these manipulations on arbovirus transmission. Virologists here and at other institutions are testing new antiviral vaccines and drugs in animal models and need tools that facilitate virus exposure of animals by the natural route of infection. We think that ATS's technology coupled with fluorescent and bioluminescent technology can be an important addition in our arsenal of tools that allow us to observe (in real time) arbovirus transmission. This approach will have an added benefit of reducing the numbers of animals that are needed for transmission and pathogenesis analyses and in evaluating antiviral protection in animals. The specific aims of this grant renewal are as follows: 1) use ATS's to develop mosquito transmission models for alphaviruses, 2) use these transmission models to understand and improve ATS's induction of RNA interference (RNAi) in mosquitoes and its impact on transmission, and 3) generate novel mosquito reporters of alphavirus infection and transmission. PUBLIC HEALTH RELEVANCE Mosquito-borne viral diseases are significant causes of illness and death throughout much of the world. We are only now beginning to understand how virus and mosquito genetics influence vector infection and virus transmission to a host. In this proposal we will develop new alphavirus-based tools to help us understand the effects of these virus and mosquito genetic factors on infection and transmission. We will develop new methods to track virus transmission from mosquitoes to experimental animals in such a way that the entire transmission cycle may be directly visualized. In this study, we plan to use fluorescent and bioluminescent technology to readily observe and detect alphavirus transmission from mosquitoes to small animals. This grant proposal will use alphavirus transducing systems (ATS's) to develop vector transmission models, identify specific alphavirus/mosquito interactions affecting virus transmission, improve ATS's ability to knock down targeted genes, and develop novel ATS-based biomarkers for mosquito infection.

描述（由申请人提供）：蚊子传播的病毒疾病仍然是世界上许多地方发病和死亡的重大原因，并对人类和动物护理基础设施造成了巨大负担。至关重要的是，我们对蚊子的载体能力和arbovirus传播的分子基础有更好的了解。基因组序列信息和其他生物信息学可用于三种主要载体物种（Anopheles Gambiae，Aedes aegypti和Culex Pipiens）。媒介生物学家和杂种动物学家仍然需要新技术，以充分利用这种信息的爆炸，并对蚊子传播的传播有了更完整的了解。在过去的15年中，艾滋病研究人员一直处于最前沿的发展，开发了被称为α型转导系统或ATS的分子工具，这些工具允许在各种蚊子（和昆虫）物种中表达基因的表达。 ATS促进了成年蚊子中的基因表达，允许对载体和α病毒基因进行功能分析，并已用于蚊子 - 阿尔帕病毒相互作用的研究。 AIDL和其他地方的媒介生物学家目前正在操纵蚊子中的先天免疫途径，并且需要ATS等工具，以表征这些操纵对Arbovirus传播的影响。这里和其他机构的病毒学家正在测试动物模型中的新抗病毒疫苗和药物，并且需要促进通过自然感染途径促进动物病毒暴露的工具。我们认为，ATS的技术以及荧光和生物发光技术可能是我们工具的重要补充，使我们可以实时观察Arbovirus的传播。这种方法将具有减少传播和发病机理所需的动物数量以及评估动物抗病毒药物所需的动物数量的额外好处。该赠款更新的具体目的如下：1）使用ATS开发用于α病毒的蚊子变速箱模型，2）使用这些传输模型来理解和改善ATS在蚊子中诱导RNA干扰（RNAI）及其对传输的影响，以及3）生成了新型的Noge Mosquito Mosquito Infressionter和Transpormation and Transpersction and Transprotection and Transperction和Transpersection。公共卫生相关性蚊子传播的病毒疾病是世界上许多地方造成疾病和死亡的重要原因。我们直到现在才开始了解病毒和蚊子遗传学如何影响载体感染和病毒传播给宿主。在此提案中，我们将开发新的基于α病毒的工具，以帮助我们了解这些病毒和蚊子遗传因素对感染和传播的影响。我们将开发新的方法，以直接可视化整个传输周期的方式来跟踪从蚊子到实验动物的病毒传播。在这项研究中，我们计划使用荧光和生物发光技术来容易观察和检测从蚊子到小动物的α病毒的传播。该赠款提案将使用α病毒转导系统（ATS）来开发向量传输模型，确定影响病毒传播的特定α病毒/蚊子相互作用，提高ATS击倒靶向基因的能力，并为蚊子感染开发基于ATS的新型生物标志物。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genetic determinants of Sindbis virus mosquito infection are associated with a highly conserved alphavirus and flavivirus envelope sequence.

辛德比斯病毒蚊子感染的遗传决定因素与高度保守的甲病毒和黄病毒包膜序列有关。

DOI：
10.1128/jvi.02060-07
发表时间：
2008
期刊：
Journal of virology
影响因子：
5.4
作者：
Pierro,DennisJ;Powers,ErikL;Olson,KenE
通讯作者：
Olson,KenE

Venezuelan and western equine encephalitis virus E1 liposome antigen nucleic acid complexes protect mice from lethal challenge with multiple alphaviruses.

委内瑞拉和西方马脑炎病毒 E1 脂质体抗原核酸复合物可保护小鼠免受多种甲病毒的致命攻击。

DOI：
10.1016/j.virol.2016.08.023
发表时间：
2016
期刊：
Virology
影响因子：
3.7
作者：
Rico,AmberB;Phillips,AaronT;Schountz,Tony;Jarvis,DonaldL;Tjalkens,RonaldB;Powers,AnnM;Olson,KenE
通讯作者：
Olson,KenE

Modeling dynamic introduction of Chikungunya virus in the United States.