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。免疫功能正常的个体是易感的，但这里的症状是自限性的。隐孢子虫也是众所周知的导致幼儿腹泻。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重视。在轮状病毒之后，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整体可处理性差。该项目的目标是为这种寄生虫开发遗传工具，这项工作建立在我们最近在实现C.小的具体来说，我们将使用手头的测定来迭代优化隐孢子虫的转染载体和方案。我们将设计策略，从受感染的动物中选择和分离稳定的转基因寄生虫，我们将使用这些工具建立隐孢子虫的报告模型。这种报告病原体将立即打开疾病，以研究和建立感染、发病机理、药物治疗和免疫保护的稳健和定量相关性。从长远来看，修改寄生虫基因组的能力将使基础发现成为可能，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.