Vesicular Trafficking of RNA from the Soma to the Germline

项目总结/摘要 动物进化出了复杂的系统，使它们的数百万个细胞能够相互交流， 其他.这些系统使细胞和组织能够在发育过程中协调行动， 改变环境条件。一些细胞与细胞之间的通讯被认为是由细胞外基质介导的。 囊泡（EV），在动物循环系统中发现的磷脂膜封闭的囊泡。一些电动汽车 含有RNA，这些EV被称为RNA EV。包封在EV内的RNA可以在生物学上是 活跃例如，EV可以将mRNA从一个组织培养细胞运输到其他组织培养细胞，并且一旦 在内化后，这些mRNA可以被翻译。因此，RNA EV可能是细胞间相互作用的重要介质。 动物的沟通 双链RNA介导的基因沉默（RNA interference，RNAi）在线虫C.优雅： 在一个体细胞中表达的dsRNA可以移动到其他体细胞。有趣的是，RNAi也是遗传的， C.线虫：dsRNA可以从体细胞运输到生殖细胞（称为RNAi遗传）。RSD-3/克林特 是一种保守的膜运输因子，被认为直接有助于囊泡的形成， 哺乳动物In C.在线虫中，我已经证明RSD-3是沉默RNA从体细胞中移动所必需的。 细胞到生殖细胞。我还发现RSD-3包被胞内囊泡， 细胞并沉积在生殖系中。我假设RSD-3 EV将RNA有效载荷从索马递送到 在正常的生殖过程中。我提出的实验是为了测试这个模型， 鉴定体内细胞间运输的内源性RNA，并探索RNA EV在细胞间运输的机制。 形成和动员。 我的研究很重要，因为它探索了两个鲜为人知的生物过程：基于RNA的 细胞间通讯和RNA指导的表观遗传。RNA EV明显存在于 大多数动物的循环液，但它们的体内功能，如果有的话，是未知的。我开发了一个系统 在动物模型中研究RNA EV的方式和原因。到目前为止，我已经确定了两个因素可能需要 RNA EV功能在C.优雅这两个因子在哺乳动物中都是保守的，暗示我在C。 线虫可以帮助我们理解RNA在所有动物细胞之间移动的方式和原因。因此，我的工作 探索RNA EV是如何产生的以及它们在体内的功能可能会提示临床上 操纵RNA EV以干预人类疾病。

PROJECT SUMMARY / ABSTRACT Animals have evolved sophisticated systems that allow their many millions of cells to communicate with each other. These systems enable cells and tissues to act in coordination during development and in response to changing environmental conditions. Some cell-to-cell communication is thought to be mediated by extracellular vesicles (EVs), phospholipid membrane-enclosed vesicles found in animal circulatory systems. Some EVs contain RNA and these EVs are referred to as RNA EVs. RNA encapsulated within EVs can be biologically active. For instance, EVs can traffic mRNAs from one tissue culture cell to other tissue culture cells and, once internalized, these mRNAs can be translated. Thus, RNA EVs may be important mediators of cell-cell communication in animals. dsRNA-mediated gene silencing (termed RNA interference, RNAi) is systemic in the nematode C. elegans: dsRNAs expressed in one somatic cell can move to other somatic cells. Interestingly, RNAi is also heritable in C. elegans: dsRNAs can be trafficked from somatic cells to germ cells (termed RNAi inheritance). RSD-3/CLINT is a conserved membrane trafficking factor, which is thought to contribute directly to vesicle formation in mammals. In C. elegans, I have shown that RSD-3 is required for the movement of silencing RNAs from somatic cells to germ cells. I have also shown that RSD-3 coats intracellular vesicles that are exocytosed from somatic cells and deposited in the germline. I hypothesize that RSD-3 EVs deliver RNA payloads from the soma to the germline during the normal course of reproduction. My proposed experiments are designed to test this model, identify endogenous RNAs trafficked between cells in vivo, and explore the mechanism by which RNA EVs are formed and mobilized. My research is important because it explores two poorly understood biological processes: RNA-based intercellular communication and RNA-directed epigenetic inheritance. RNA EVs are clearly present in the circulatory fluids of most animals but their in vivo function, if any, is not known. I have developed a system to study the how and why of RNA EVs in an animal model. To date, I have identified two factors likely required for RNA EV function in C. elegans. Both of these factors are conserved in mammals, hinting that my findings in C. elegans may help us understand how and why RNAs move between cells in all animals. Therefore, my work exploring how RNA EVs are produced and what function they have in vivo may suggest ways to clinically manipulate RNA EVs in order to intervene in human disease.