Yolk protein-mediated germline delivery in vector snails

总结 基于遗传的病媒控制在防治病媒传播疾病方面大有希望。这样一个充满希望 然而，这一战略尚未在任何血吸虫病媒介钉螺控制项目中得到检验， 世界上最具破坏性的寄生虫病。关键原因是缺乏生殖细胞输送技术， 将遗传物质送入配子或胚胎。无法将遗传物质输送到蜗牛体内 生殖系细胞是由于其配子的复杂解剖结构，其卵子的极小尺寸， 和它不寻常的生殖系统，这排除了所有现有的，发达的遗传应用， 交付技术。为了规避这些限制，我们提出了一种创新的方法，利用蜗牛 卵黄蛋白作为载体，通过受体的自然过程将遗传物质穿梭到生长的卵母细胞中， 在卵黄发生过程中介导的内吞作用。两种最有前途的蜗牛卵黄蛋白，卵黄铁蛋白 和卵黄蛋白原，两者都存在于血吸虫病媒介蜗牛光滑双脐螺的基因组中， 将被调查。我们将首先从蜗牛卵黄铁蛋白中鉴定出一种高效的卵母细胞受体结合配体 或卵黄蛋白原（Aim 1），然后测试所鉴定的配体在递送成簇的规则间隔的短 在一些实施方案中，CRISPR载体可以将回文重复序列（CRISPR）货物引入卵母细胞中用于种系修饰（Aim 2）。这项研究将开启一个 为血吸虫病基础和应用研究的所有遗传操作研究打开了一扇令人兴奋的大门。

SUMMARY Genetics-based vector control holds great promise in combating vector transmitted diseases. Such a promising strategy, however, has not yet been tested in any vector snail control program of schistosomiasis, one of the world’s most devastating parasitic diseases. The key reason is the lack of a germline delivery technology for delivering genetic materials into gametes or embryos. The inability to deliver genetic materials into snail germline cells is due to the complex anatomical structure of its gametes, the exceedingly small size of its eggs, and its unusual reproductive system, which preclude the application of all existing, well-developed genetic delivery technologies. To circumvent these limitations, we propose an innovative approach that exploits snail yolk proteins as vehicles for shuttling genetic materials into growing oocytes via the natural process of receptor- mediated endocytosis during vitellogenesis. Two types of the most promising snail yolk proteins, yolk ferritin and vitellogenin, both found present in the genome of the schistosomiasis vector snail Biomphalaria glabrata, will be investigated. We will first identify a highly efficient oocyte receptor-binding ligand from snail yolk ferritin or vitellogenin (Aim1) and then test the identified ligand in delivering clustered regularly interspaced short palindromic repeats (CRISPR) cargo into the oocytes for germline modification (Aim 2). This study will open an exciting door for all genetic manipulation studies for basic and applied research of schistosomiasis.