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

• 批准号: 8302208
• 负责人: JINSONG ZHU
• 金额: $ 19.95万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8302208
• 关键词: 3' 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

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.

描述（由申请人提供）：2009年全球报告了2.43亿个疟疾病例和863,000例死亡。要从一个人传播到另一个人，疟疾寄生虫（Plasmodium spp。）必须完成蚊子的精致发展计划并生存。蚊子先天免疫系统的攻击。我们的长期目标是发现参与抗质量防御的蚊子microRNA，阐明这些microRNA的分子功能，并利用这些知识来帮助未来的努力控制疟疾传播。 在大多数真核生物中，microRNA是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，并与特定靶标mRNA的3'未翻译区域相关，以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，一些microRNA的水平在中肠受到疟原虫感染显着影响。 RNAi介导的DICER-1或ARGONAUTE-1（microRNA途径的关键成分）的沉默已被证明可以增强寄生虫存活。因此，我们假设microRNA在调节对疟疾寄生虫的蚊子防御反应中起着重要作用。 为了检验这一假设，我们需要找到仅在蚊子暴露于血液粉中疟疾寄生虫后，这些蚊子mRNA受microRNA的特异性调节。仅通过计算方法的microRNA目标预测通常会受到高假阳性速率的阻碍。我们将使用hits-clip（通过交联免疫沉淀分离的RNA的高通量测序）代替捕获和识别mirisc中Argonaute-1绑定的microRNA和靶标mRNA。这将揭示中肠中所有信使RNA中的精确目标站点。该项目的具体目的是：（1）确定恶性疟原虫入侵疟原虫在富含Mirisc的冈比亚巨型MicroRNA。 （2）识别蚊子信使RNA因质量挑战而被microRNA选择性地调节。 我们提出的研究涉及对蚊子相互作用中microRNA功能的严重差距，并可能为阻断疟疾传播提供新的分子靶标。