Transport of gene silencing between cells during RNA interference

项目摘要/摘要 RNA干扰(RNAi)是双链RNA特异性失活的一种保守机制 序列匹配的基因。RNAi从诺贝尔奖获得者在简单的 蠕虫线虫寻求一种治疗方法来沉默缺乏常规致病基因的人类致病基因 药物。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治疗。模式生物研究的持续相关性和意义 秀丽线虫的突出特点是，将RNA输入线虫的RNA通道的人类对应物。 RNAi过程中的线虫细胞是将基于RNAi的药物导入人类细胞所必需的。 应聘者提交了一份为期5年的职业发展计划，旨在利用线虫获得基础知识 洞察RNA干扰过程中细胞之间的RNA运输，同时建立一个独立的 在研究型大学的学术生涯。候选人将建立在他雄厚的遗传学和 生物化学将在克雷格博士的指导下发展成为RNA运输的独立研究人员 亨特是研究动物细胞间RNA运输的先驱和领先者。该计划将在#年实施。 哈佛大学分子生物学系，现代生物学的领先机构。 导师实验室的研究导致发现了保守的RNA通道SID-1，这是 RNAi介导的沉默信号的输入和组织内细胞之间的信号传输。在最近一次 发表后，候选人报告发现，线虫组织的RNA输出是通过一种 调节SID-1的非依赖机制。在指导阶段，候选人将：1)剖析SID-1 使用先进的显微镜观察RNA在组织内细胞之间的依赖运输，并研究 SID-1从组织中独立输出RNA是由SID-3介导的，SID-3是这种输出所需的基因； 研究细胞内RNAi途径蛋白在产生从该细胞运输的RNAs中的作用。在……里面 除了导师的实验室外，Aim1的高级显微镜将在 候选人的合作者庄晓伟博士，他是超分辨率显微镜的先驱。在.期间 在奖项的独立阶段，候选人将分析另外两个基因Sexd-1和Sexd-2的作用 由候选人发现的，是组织间输出所必需的，并将定义基本的分子途径 使用在指导阶段获得的方法和技术进行导出 培训补充细胞生物学、遗传学和生物化学方法，同时执行上述 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作为 独立调查员。此外，拟议的研究将揭示rna运输的基本方面。 在线虫的RNAi过程中，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.