Experimental identification of microRNA targets in mosquitoes during Plasmodium i

蚊子疟原虫 i 期 microRNA 靶标的实验鉴定

基本信息

批准号：
8095042
负责人：
JINSONG ZHU
金额：
$ 23.64万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-15 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8095042
关键词：
3&apos Untranslated Regions Address Affect Animals Anopheles Genus Anopheles gambiae Antibodies Binding Binding Sites Bioinformatics Biological Assay Bite Blood Brain Caenorhabditis elegans Cessation of life Code Competence Complex Culicidae Data Set Development Epithelium Eukaryota Female Future Gene Expression Gene Expression Profile Gene Expression Regulation Genome Goals Human Immune Immune system Immunoprecipitation Infectious Agent Ingestion Knowledge Lead Learning Luciferases Malaria Maps Mediating Messenger RNA Methods MicroRNAs Midgut Molecular Molecular Target Mus Nucleotides Oocysts Parasites Pathway interactions Persons Physiology Plants Plasmodium Plasmodium berghei Plasmodium falciparum Play Post-Transcriptional Regulation Process Protein Biosynthesis Proteins RNA RNA Binding RNA Interference RNA Sequences RNA-Induced Silencing Complex Reaction Reading Regulation Reporter Reporting Research Role Sampling Site Small RNA Staging Testing Time Transfection Translational Repression Translations base crosslink defense response feeding genetic manipulation human DICER1 protein improved insight mRNA Transcript Degradation mature animal novel programs protein complex response transmission process ultraviolet irradiation vector vector mosquito

项目摘要

DESCRIPTION (provided by applicant): 243 million malaria cases and 863,000 attributed deaths were reported globally in 2009. To be transmitted from one person to another, the malaria parasite (Plasmodium spp.) has to complete an elaborate developmental program in the mosquito and survive the attacks from mosquito innate immune system. Our long term goal is to discover mosquito microRNAs that are involved in anti-Plasmodium defense, elucidate the molecular functions of these microRNAs, and to use this knowledge to aid future efforts to control malaria transmission. MicroRNAs are small endogenous regulatory molecules in most eukaryotes. In animals, mature microRNAs are integrated into a microRNA-induced silencing complex (miRISC) and associated with the 3' untranslated regions of specific target mRNAs to suppress gene expression either through inhibition of translation or through mRNA degradation. In Anopheles gambiae mosquitoes, levels of some microRNAs are markedly affected in the midgut by Plasmodium infection. RNAi-mediated silencing of either Dicer-1 or Argonaute-1, key components of the microRNA pathway, has been shown to enhance parasite survival. We therefore hypothesize that microRNAs play an important role in modulating mosquito defense response to malaria parasites. To test this hypothesis, we need to find mosquito mRNAs that are specifically regulated by microRNAs only after mosquitoes are exposed to malaria parasites in a blood meal. microRNA target prediction by computational approaches alone is generally hampered by high false-positive rates. We will use the HITS-CLIP (high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation) method instead to capture and identify both the microRNAs and target mRNAs bound by Argonaute-1 in miRISC. This will reveal precise target sites across all messenger RNAs expressed in the midgut. The specific aims of this project are to: (1) Determine Anopheles gambiae microRNAs that are enriched in miRISC during Plasmodium falciparum invasion. (2) Identify mosquito messenger RNAs that are selectively regulated by microRNAs in response to Plasmodium challenge. Our proposed studies address a serious gap in the understanding of microRNA functions in the mosquito-Plasmodium interactions, and may provide novel molecular targets for blocking malaria transmission. PUBLIC HEALTH RELEVANCE: The female Anopheles mosquito is the vector for human malaria. Some small RNA molecules in the mosquito are implicated in regulating the mosquito defense reactions against malaria parasite. Our research will identify the targets of those small RNAs and the insights in turn may lead to new strategies for malaria control.

描述（由申请人提供）：2009 年，全球报告了 2.43 亿例疟疾病例和 863,000 例死亡病例。为了从一个人传播到另一个人，疟疾寄生虫（疟原虫属）必须在蚊子体内完成复杂的发育程序并存活下来来自蚊子先天免疫系统的攻击。我们的长期目标是发现参与抗疟原虫防御的蚊子 microRNA，阐明这些 microRNA 的分子功能，并利用这些知识来帮助未来控制疟疾传播。 MicroRNA 是大多数真核生物中小的内源性调节分子。在动物中，成熟的 microRNA 被整合到 microRNA 诱导的沉默复合物 (miRISC) 中，并与特定目标 mRNA 的 3' 非翻译区相关，通过抑制翻译或通过 mRNA 降解来抑制基因表达。在冈比亚按蚊中，中肠中一些 microRNA 的水平受到疟原虫感染的显着影响。 RNAi 介导的 Dicer-1 或 Argonaute-1（microRNA 途径的关键组成部分）的沉默已被证明可以增强寄生虫的存活率。因此，我们假设 microRNA 在调节蚊子对疟疾寄生虫的防御反应中发挥着重要作用。为了检验这一假设，我们需要找到仅在蚊子接触血液中的疟疾寄生虫后才受到 microRNA 特异性调节的蚊子 mRNA。仅通过计算方法进行 microRNA 靶标预测通常会受到高假阳性率的阻碍。我们将使用 HITS-CLIP（通过交联免疫沉淀分离的 RNA 高通量测序）方法来捕获和识别 miRISC 中 Argonaute-1 结合的 microRNA 和目标 mRNA。这将揭示中肠中表达的所有信使 RNA 的精确靶位点。该项目的具体目标是：（1）确定冈比亚按蚊在恶性疟原虫入侵期间富含miRISC的microRNA。 (2) 鉴定蚊子信使 RNA，这些 RNA 受 microRNA 选择性调节，以响应疟原虫的攻击。我们提出的研究解决了对蚊子与疟原虫相互作用中 microRNA 功能的理解上的严重差距，并可能为阻止疟疾传播提供新的分子靶点。公共卫生相关性：雌性按蚊是人类疟疾的传播媒介。蚊子体内的一些小 RNA 分子参与调节蚊子针对疟疾寄生虫的防御反应。我们的研究将确定这些小 RNA 的靶标，这些见解反过来可能会带来控制疟疾的新策略。