Genetic analysis of Cryptosporidium

描述（由申请人提供）：隐孢子虫在HIV-AIDS患者中引起危及生命的肠道疾病。免疫能力强的个体易受感染，但症状是自限性的。众所周知，隐孢子虫也会引起幼儿腹泻。然而，隐孢子虫对儿童死亡率和发病率的全面影响直到最近才得到认识。继轮状病毒之后，隐孢子虫是导致2岁以下儿童严重腹泻的最常见病原体。实现迫切需要的进展的主要障碍是隐孢子虫在实验室中的总体难处理性。这个项目的目标是开发这种寄生虫的遗传工具，这项工作建立在我们最近在实现小疟原虫转染方面的突破之上。具体来说，我们将使用手头的分析迭代优化隐孢子虫的转染载体和方案。我们将制定策略，从感染动物中选择和分离稳定的转基因寄生虫，并将利用这些工具建立隐孢子虫的报告模型。这些报告病原体将立即开启疾病的研究，并建立与感染、发病机制、药物治疗和免疫保护相关的强有力的定量关系。从长远来看，修改寄生虫基因组的能力将使根本性的发现成为可能，并转化为治疗和预防隐孢子虫病的工具和机会。

DESCRIPTION (provided by applicant): Cryptosporidium causes life threatening enteric disease in HIV-AIDS patients. Immunocompetent individuals are susceptible, but here symptoms are self-limiting. Cryptosporidium is also well known to cause diarrhea in small children. However, the full impact of Cryptosporidium on child mortality and morbidity was only recently appreciated. After rotavirus, Cryptosporidium is the most common pathogen responsible for severe diarrhea in children under 2 years. The main roadblock to achieving urgently needed advances is the overall poor tractability of Cryptosporidium in the laboratory. The goal of this project is to develop genetic tools for this parasite, and the effort builds on ou recent breakthrough in achieving transfection for C. parvum. Specifically, we will use the assay in hand to iteratively optimize transfection vectors and protocols for Cryptosporidium. We will devise strategies to select and isolate stable transgenic parasites from infected animals, and we will use these tools to establish a reporter model for Cryptosporidium. Such reporter pathogens will immediately open the disease to study and establish robust and quantitative correlates of infection, pathogenesis, drug cure, and immune-protection. In the long term, the ability to modify the parasite's genome will enable fundamental discovery and translate into tools and opportunities to treat and prevent cryptosporidiosis.