Development of genetic tools for the study of mosquito hemocytes

项目总结 蚊子已经形成了一种强大的天然免疫系统，包括细胞免疫和体液免疫 共同塑造蚊子媒介能力的反应，因此引起了极大的兴趣 作为潜在的目标，使蚊子不易传播疾病。免疫细胞，称为血细胞， 是蚊子细胞和体液免疫反应的中心，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，尽管它们很重要，但对 由于缺乏遗传资源，蚊子血细胞一直受到限制，这使得它们的特征仅限于 大小和形状的形态特征。有一个长期目标是描述蚊子免疫细胞的特征 种群及其对先天免疫的贡献，这一应用的总体目标是开发 遗传工具和资源，以提高我们对蚊子免疫细胞功能的了解。最近的单电池 研究已经用新的和提高的分辨率表征了蚊子免疫细胞群，提供了 确定定义免疫细胞亚群的候选通用或亚型特异性标记基因。 作为这一提议的基础和理论基础，这些候选血细胞的调节区 标记基因将被用来产生驱动荧光标记的转基因按蚊(目标1) 或转录激活因子(AIM 2)在不同的免疫细胞群中。总之，我们提议的实验将 为蚊子血细胞群落提供有价值的新遗传资源，这将显著促进我们的 了解按蚊免疫细胞生物学，同时为今后更好的研究奠定基础 确定血细胞对疟疾寄生虫杀灭的贡献。

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.