The role of mosquito microRNAs in blood digestion.

蚊子 microRNA 在血液消化中的作用。

• 批准号: 9067816
• 负责人: ALEXANDER SIMEON RAIKHEL
• 金额: $ 41.54万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9067816
• 关键词: 3' Untranslated Regions; Aedes; Anopheles gambiae; Antibodies; Binding Sites; Blood; Cell Line; Computational algorithm; Computer Analysis; Culicidae; Data; Digestion; Enzymes; Evaluation; Event; Female; Gene Expression; Gene Proteins; Gene Targeting; Genes; Health; Human; Immunity; Knowledge; Link; Luciferases; MicroRNAs; Midgut; Molecular; Orthologous Gene; Peptide Hydrolases; Phenotype; Physiological; Play; Porifera; Process; Proteins; RNA; RNA Interference; Regulation; Reporter; Reproduction; Reproductive Process; Role; Specificity; Testing; Transfection; Transgenic Organisms; Untranslated RNA; Untranslated Regions; Vector-transmitted infectious disease; base; feeding; gene interaction; human disease; in vitro Assay; in vitro testing; in vivo; innovation; knock-down; loss of function; pathogen; programs; spatiotemporal; vector control; vector mosquito

DESCRIPTION (provided by applicant): Female mosquitoes require a blood meal for reproduction, and during blood feeding spread many of the devastating vector-borne diseases in humans. A deeper understanding of the molecular mechanisms linked to mosquito blood meal processes, reproductive events and anti-pathogen immunity is of particular importance for devising innovative vector control strategies. Based on strong preliminary data, we suggest that microRNAs (miRNA) play important and conserved roles in blood digestive functions of vector mosquitoes. We have developed antagomirs for depletion of miRNAs and their functional characterization in Aedes aegypti and Anopheles gambiae. We have identified two miRNAs with strong effects on blood digestion. Evaluation of spatiotemporal specificity of these miRNAs will be achieved by utilizing GAL4-UAS miRNA Tough Decoy or sponge loss-of-function transgenic mosquitoes. An important step in understanding miRNA function is determining their targets. A multi-algorithm computational target-prediction approach will be used for determination of the miRNA targets. Luciferase reporter constructs carrying 3 2UTRs of the candidate miRNA targets will then be transfected into mosquito cell lines, and their expression levels will be tested after ectopic co-transfection of a miRNA that is being studied. A phenotype rescue will be achieved by RNA interference knockdown of a target gene in the transgenic loss-of-function miRNA mosquitoes. As an alternative approach, we will identify miRNAs complementary to 32UTRs of genes encoding key enzymes involved in blood digestion and study their respective roles. This application presents a comprehensive approach for functional characterization of miRNAs in two of the most important mosquito vectors for human diseases.

描述（由申请人提供）：雌性蚊子需要吸血来繁殖，在吸血过程中会在人类中传播许多毁灭性的媒介传播疾病。更深入地了解与蚊子血粉过程、繁殖事件和抗病原体免疫相关的分子机制对于设计创新的病媒控制策略特别重要。基于强有力的初步数据，我们认为 microRNA (miRNA) 在媒介蚊子的血液消化功能中发挥着重要且保守的作用。我们开发了 antagomir，用于消除埃及伊蚊和冈比亚按蚊中的 miRNA 及其功能表征。我们已经鉴定出两种对血液消化具有强烈影响的 miRNA。这些 miRNA 的时空特异性评估将通过利用 GAL4-UAS miRNA Tough Decoy 或海绵功能丧失转基因蚊子来实现。了解 miRNA 功能的一个重要步骤是确定其靶点。多算法计算目标预测方法将用于确定 miRNA 目标。然后将携带候选 miRNA 靶标的 3 个 2UTR 的荧光素酶报告基因构建体转染到蚊子细胞系中，并在之后测试它们的表达水平 正在研究的 miRNA 的异位共转染。通过 RNA 干扰敲除转基因功能丧失 miRNA 蚊子中的目标基因，可以实现表型拯救。作为一种替代方法，我们将鉴定与编码参与血液消化的关键酶的基因的 32UTR 互补的 miRNA，并研究它们各自的作用。该应用提出了一种综合方法，用于对人类疾病的两种最重要的蚊子媒介中的 miRNA 进行功能表征。