Generation of transmission-compromised mosquitoes

传播受限的蚊子的产生

基本信息

批准号：
10039237
负责人：
Deborah J Andrew
金额：
$ 24.56万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-20 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10039237
关键词：
Adult Aedes Affect African Trypanosomiasis Age Anopheles gambiae Automobile Driving Biological Assay Blood CRISPR/Cas technology Candidate Disease Gene Cause of Death Cells Cessation of life Child Clinical Communicable Diseases Culicidae Dengue Dengue Fever Dengue Hemorrhagic Fever Dengue Virus Disease Distant Drops Drosophila genus Event Expression Profiling Female Fertility Generations Genes Goals Head Homeostasis Human Incidence Individual Insect Vectors Insecta Invaded Laboratories Learning Leishmaniasis Life Cycle Stages Link Longevity Lyme Disease Maintenance Malaria Morphogenesis Morphology Mosquito-borne infectious disease Mutation Ocular Onchocerciasis Organ Outcome Parasites Pathway interactions Persons Phenotype Population Publications RNA Interference Reporting Role Saliva Salivary Glands Specific qualifier value Sporozoites Testing Time Tissues Tube Virus Work ZIKA base conditional knockout cost disease transmission feeding fitness genetic testing gland development human disease knock-down malaria infection pathogen prevent transcription factor transmission process vector mosquito

项目摘要

Insect-borne diseases kill more humans than all other diseases combined. Existing strategies for limiting the spread of such diseases are helping, but it is clear that complete eradication of some of the most deadly diseases – especially malaria – is going to require additional approaches. The goal of this project is to test strategies to block insect-borne disease transmission by intervening at the level of the insect vector, preventing transmission of disease-causing parasites and viruses by causing the timely death of the adult female mosquito salivary gland (SG). A current limitation for studies attempting to manipulate the SG – a gateway organ for transmitting malaria and other insect borne diseases - is that almost nothing is known about SG development, function or homeostasis in any insect outside Drosophila. The molecules involved in specifying, building or specializing mosquito SGs are largely unknown. The proposed study begins with learning the consequences of RNAi knock-down of three Anopheles gambiae transcription factor (TF) genes whose Drosophila orthologues have key roles in Drosophila SG development, function and survival. We ask if TF gene knockdown affects SG survival or morphology, or the ability of infective malaria sporozoites to invade the SGs and to be transmitted in the saliva. In the second aim, we create null and tissue-specific mutations in the Anopheles gambiae sage gene, which encodes a SG-specific TF whose Drosophila orthologue is required for SG survival and expression of SG-specific cargo genes. We ask how complete loss of sage affects viability, fecundity and SG morphology. We ask if loss of sage affects sporozoite SG invasion and sporozoite counts in saliva. We also determine if SG-specific loss of sage following a blood meal leads to similar outcomes. Finally, to learn how well our SG-centric approach to limiting disease transmission works in general, we ask if RNAi knockdown of the sage orthologue in Aedes aegypti affects the SG and its ability to transmit Dengue virus.

昆虫传播的疾病导致的死亡人数比所有其他疾病的死亡人数总和还多。此类疾病的传播正在有所帮助，但很明显，彻底根除一些最致命的疾病疾病——尤其是疟疾——将需要额外的方法，该项目的目标是进行测试。通过在昆虫媒介水平上进行干预来阻止虫媒疾病传播的策略，预防通过导致成年女性及时死亡来传播致病寄生虫和病毒蚊子唾液腺（SG）是目前试图操纵 SG 的研究的局限性——一个门户。传播疟疾和其他昆虫传播疾病的器官 - 人们对 SG 几乎一无所知果蝇以外任何昆虫的发育、功能或稳态涉及指定的分子，构建或专门的蚊子 SG 在很大程度上是未知的。拟议的研究从了解蚊子 SG 开始。 RNAi 敲除冈比亚按蚊的三个转录因子 (TF) 基因的后果果蝇直系同源物在果蝇 SG 的发育、功能和生存中发挥着关键作用。基因敲低会影响 SG 的存活或形态，或感染性疟疾子孢子侵入宿主细胞的能力。 SG 并在唾液中传播在第二个目标中，我们在唾液中创建无效突变和组织特异性突变。冈比亚按蚊鼠尾草基因，编码 SG 特异性 TF，其果蝇直系同源物是 SG 的存活和 SG 特异性货物基因的表达我们询问鼠尾草的完全丧失如何影响生存能力，我们询问鼠尾草的损失是否会影响子孢子的 SG 入侵和子孢子计数。我们还确定了血粉后 SG 特异性的鼠尾草损失是否会导致类似的结果。最后，为了了解我们以 SG 为中心的方法来限制疾病传播的总体效果如何，我们询问是否埃及伊蚊中鼠尾草直系同源物的 RNAi 敲低会影响 SG 及其传播登革热的能力病毒。