Development of genetic tools for the study of mosquito hemocytes

项目摘要 蚊子已经形成了一个强大的先天免疫系统，包括细胞免疫和体液免疫。 这些反应共同塑造了蚊子的媒介能力，因此引起了人们的极大兴趣。 作为潜在的目标，使蚊子难以传播疾病。免疫细胞，也就是血细胞， 是蚊子细胞和体液免疫反应的核心，包括吞噬作用、建立 免疫记忆，以及识别和杀死疟疾寄生虫。然而，尽管他们的重要性，研究 蚊子血细胞由于缺乏遗传资源而受到限制， 尺寸和形状的形态学特性。长期目标是表征蚊子免疫细胞 由于本申请的总体目标是研究人类群体及其对先天免疫的贡献， 遗传工具和资源，以提高我们对蚊子免疫细胞功能的了解。最近单细胞 研究已经以新的和增加的分辨率表征了蚊子免疫细胞群体， 鉴定定义免疫细胞亚群的候选通用或亚型特异性标记基因。 作为该提议的基础和理论基础，这些候选血细胞的调节区域 标记基因将用于产生驱动荧光标记的转基因按蚊（Aim 1） 或转录激活因子（Aim 2）。总之，我们提出的实验将 为蚊子血细胞群落提供宝贵的新遗传资源，这将大大促进我们的 了解按蚊的免疫细胞生物学，同时作为未来研究的基础，以更好地 定义血细胞对杀死疟原虫的贡献。

PROJECT SUMMARY Mosquitoes have developed a robust innate immune system comprised of cellular and humoral immune responses that together shape mosquito vector competence, which as a result has garnered significant interest as potential targets to make mosquitoes refractory to disease transmission. Immune cells, known as hemocytes, are central to mosquito cellular and humoral immune responses that include phagocytosis, the establishment of immune memory, and the recognition and killing of malaria parasites. Yet, despite their importance, studies of mosquito hemocytes have been limited by a lack of genetic resources that have limited their characterization to morphological properties of size and shape. With a long-term goal to characterize mosquito immune cell populations and their contributions to innate immunity, the overall objective of this application is to develop genetic tools and resources to improve our understanding of mosquito immune cell function. Recent single-cell studies have characterized mosquito immune cell populations with new and increased resolution, providing the identification of candidate universal or subtype-specific marker genes that define immune cell sub-populations. Serving as the foundation and rationale of this proposal, regulatory regions of these candidate hemocyte marker genes will be used to generate transgenic Anopheles mosquitoes that drive fluorescent markers (Aim 1) or transcriptional activators (Aim 2) in distinct immune cell populations. Together, our proposed experiments will provide valuable new genetic resources for the mosquito hemocyte community that will significantly advance our understanding of Anopheles immune cell biology, while serving as the foundation for future studies to better define the contributions of hemocytes to malaria parasite killing.