Transport of gene silencing between cells during RNA interference

项目总结/摘要 RNA干扰（RNA interference，RNAi）是一种保守的机制，通过它双链RNA可以特异性地干扰RNA， 匹配序列的基因。RNAi已经从一个简单的获得诺贝尔奖的发现迅速发展起来， 沃姆角一种治疗方法，以沉默人类致病基因，缺乏传统的 药然而，RNAi的基本方面仍然不清楚，需要了解它们以确保 安全有效的RNAi疗法。模式生物研究的持续相关性和意义 C. elegans是强调的事实，即人类对应的RNA通道，进口RNA到C。 在RNAi过程中，线虫细胞是将基于RNAi的药物导入人类细胞所必需的。 候选人提出了一个5年的职业发展计划，旨在使用C。优雅的获得基本的 深入了解RNA干扰过程中细胞间RNA的转运，同时建立一个独立的 研究型大学的学术生涯。候选人将建立在他在遗传学方面的坚实基础上， 在克雷格博士的指导下， Hunter是动物细胞间RNA转运研究的先驱和领导者。该计划将于 哈佛大学的分子生物学系，现代生物学的领先机构。 导师实验室的研究导致了保守RNA通道SID-1的发现，该通道是转录所需的。 RNAi介导的沉默信号的输入和组织内细胞之间的信号转运。在最近的一 发表，候选人报告的发现，出口的RNA从C。线虫组织通过一种 规范SID-1独立机制。在指导阶段，候选人将：1）剖析SID-1 使用先进的显微镜技术，研究组织内细胞之间RNA的依赖性转运， 来自组织的RNA的SID-1独立输出由sid-3介导，sid-3是这种输出所需的基因;和2） 研究细胞内RNAi途径蛋白在产生从该细胞转运的RNA中的作用。在 除了导师的实验室外，Aim 1的高级显微镜将在 候选人的合作者Xiaowei Zhuang博士是超分辨率显微镜的先驱。期间 在该奖项的独立阶段，候选人将分析另外两个基因sexd-1和sexd-2的作用 候选人发现了组织间输出所需的并将定义基本分子途径 使用在辅导阶段获得的方法和技术， 在执行上述工作的同时，进行补充细胞生物学、遗传学和生物化学方法的培训 研究计划将使候选人能够建立一个多方面和丰富的研究计划， 独立调查员此外，拟议的研究将揭示RNA转运的基本方面 在C. elegans，这将影响治疗人类疾病的RNAi方法的设计。

Project Summary/Abstract RNA interference (RNAi) is a conserved mechanism by which double-stranded RNA can specifically inactivate genes of matching sequence. RNAi has rapidly developed from a Nobel-prize winning discovery in the simple worm C. elegans to a therapeutic approach to silence human disease-causing genes that lack conventional medicines. However, fundamental aspects of RNAi remain unclear and they need to be understood to ensure safe and efficacious RNAi therapy. The continued relevance and significance of studies in the model organism C. elegans is underscored by the fact that the human counterpart of an RNA channel that imports RNA into C. elegans cells during RNAi is required for the import of RNAi-based drugs into human cells. The candidate presents a 5-year career development plan that aims to use C. elegans to gain fundamental insights into the transport of RNA between cells during RNA interference, while establishing an independent academic career at a research university. The candidate will build on his strong foundation in genetics and biochemistry to develop into an independent researcher in RNA transport under the mentorship of Dr. Craig Hunter, a pioneer and leader in the study of RNA transport between animal cells. The plan will be carried out in the Department of Molecular Biology at Harvard University, a leading institution in modern biology. Research in the mentor's lab led to the discovery of the conserved RNA channel SID-1 that is required for the import of RNAi-mediated silencing signals and the transport of signals between cells within a tissue. In a recent publication, the candidate reported the discovery that export of RNA from C. elegans tissues occurs through a regulated SID-1 independent mechanism. During the mentored phase, the candidate will: 1) Dissect the SID-1 dependent transport of RNA between cells within a tissue using advanced microscopy and examine how the SID-1 independent export of RNA from tissues is mediated by sid-3, a gene required for such export; and 2) examine the role of RNAi pathway proteins within a cell in generating RNAs transported from that cell. In addition to the mentor's laboratory, advanced microscopy for Aim1 will be carried out in the lab of the candidate's collaborator Dr. Xiaowei Zhuang, who is a pioneer in super-resolution microscopy. During the independent phase of the award, the candidate will analyze the roles of sexd-1 and sexd-2, two other genes discovered by the candidate that are required for inter-tissue export and will define a basic molecular pathway for export using the approaches and techniques acquired during the mentored phase Training in the complementary cell biological, genetic, and biochemical approaches while executing the above research plan will equip the candidate to establish a multi-faceted and rich research program as an independent investigator. Further, the proposed studies will reveal fundamental aspects of RNA transport during RNAi in C. elegans, which will impact the design of therapeutic RNAi approaches to human diseases.