Function and Mechanism of Coat Protein Complex I in RNA Silencing

本项目的重点是Coat Protein Complex I(COPI)在将一组酶运输到植物细胞内的正确位置以执行调节功能方面的作用，包括降解mRNA和抑制蛋白质合成。通过加强对基因沉默机制的了解，该项目可以加强生物技术方法来改良农作物。该项目还将把研究经验融入遗传学课程，以加强对本科生的基因组学教学。该项目还为高中生提供遗传学和生物学方面的实践培训。该项目的目标是阐明COPI如何促进miRNA介导的RNA沉默。MiRNAs是一组小的非编码调控RNA，抑制内源基因和侵袭性RNA的表达。MiRNAs被加载到ArgAerte蛋白质中，形成RNA诱导的沉默复合体(mi-RISC)，进而摧毁目标RNA。虽然已经确定和研究了微型生物多样性委员会的关键组成部分和辅助伙伴，但许多新的调节层或因素仍在恢复中；确切的机制仍有待开发。利用一种新的遗传筛选方法，PI发现了一个新的基因，CoPI2-COP，它编码α的一个亚基，是miRNA途径中的一个新角色。中心假说是，COPI负责将微-RISC和/或P-体的成分运送到其功能部位，以影响RNA沉默。PI将应用一系列遗传、细胞生物学和生化方法来验证这一假设。对COPI的功能和机制分析不仅将揭示RNA沉默的新调节层和机制，还将使我们能够提高RNAi技术的效率。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project focuses on the role of Coat protein complex I (COPI) in transporting a group of enzymes to the right places inside plant cells to perform regulatory functions, including degradation of mRNA and inhibition of protein synthesis. By enhancing understanding of the mechanisms of gene silencing, this project can enhance biotechnological approaches to improve agricultural crops. This project will also incorporate research experiences into a Genetics course to strengthen the teaching of genomics to undergraduates. The project also provides hands-on training for high school students in genetics and biology. The goal of this project is to elucidate how COPI contributes to miRNA-mediated RNA silencing. miRNAs are a group of small noncoding regulatory RNAs that suppress expression of endogenous genes and invasive RNAs. miRNAs are loaded into Argonaute proteins to form RNA-induced silencing complexes (mi-RISC) that in return destroy target RNAs. Although key components and auxiliary partners of mi-RISC have been identified and studied, many new regulatory layers or factors are still being recovered; and the precise mechanisms remain to be exploited. Using a novel genetic screening approach, the PI has identified a new gene, α2-COP that encodes a subunit of COPI, as a new player in the miRNA pathway. The central hypothesis is that COPI is responsible for delivering components of mi-RISC and/or P-bodies to their functional sites to impact RNA silencing. The PI will apply a series of genetic, cell biological and biochemical approaches to test this hypothesis. Functional and mechanistic analyses of COPI will not only reveal new regulatory layers and mechanisms of RNA silencing, but also enable us to improve the efficacy of RNAi technology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.