非编码RNA代谢与功能

With the advent of the RNA-sequencing and other newly developed technologies in modern Biology, it has been increasingly established that noncoding RNAs (ncRNAs) are broadly expressed in all kingdoms of animals and plants and play important roles in diverse biological processes, such as development, immune responses, oncogenesis and aging. Understanding the biogenesis, function and mechanisms of action of different classes of regulatory RNAs is becoming the central topic of the modern molecular biology. However, it still remains largely unexplored on the generation, processing and degradation of several types of recently discovered ncRNAs as well as how their processing is linked to function. Based on the recent achievements supported by the previous NSFC Major Research Plan on Regulatory RNAs (2014-2019), this integrated project containing seven research laboratories will focus on the newly discovered small inference RNAs, long noncoding RNAs and circular RNAs to dissect the underlying mechanisms that control their production, processing, structure conformation and degradation with cutting-edge methodologies in molecular and cell biology, genetics and computational biology. Functional analysis of several ncRNAs will be employed in model organisms including C. elegans, D. melanogaster, A. thaliana and mouse. Specifically, we will elucidate the production of risiRNAs (antisense ribosomal siRNAs) and how these small RNAs regulate pre-rRNA through the nuclear RNAi pathway in worms; we will understand the biogenesis and biological activity of the 22nt long secondary siRNA as a part of regulatory cascades in plants. We will continue our efforts on elucidating the biogenesis, structure and degradation of circular RNAs as well as the translational regulation of a subset of circular RNAs. These will include understanding how hundreds of circular RNAs are produced from the Discam gene in flies, identifying new nucleases that can degrade these stable RNA circles, analyzing circular RNA-protein complexes in cells and in vitro, and dissecting their structural conformation with or without protein partners. Finally, using CRISPR/Cas9-mediated genome editing, we will build up gene models to understand how eRNAs act in regulating chromatin looping and mouse models to address the functional significance of evolutionarily conserved lincRNAs. Collectively, these proposed studies will continue to explore the biogenesis and function of recently identified ncRNA species. Understanding these fundamental features of ncRNAs have the potential to provide new insights into their functional significance in different biological settings.

随着高通量测序技术和现代生物学研究手段的飞速发展和应用，非编码RNA的普遍性与重要性越来越显著，它们广泛参与人类和动植物的生命活动，包括生殖发育、免疫应答、癌症和衰老等重要生理和病理过程。解析非编码RNA的功能与作用机制已成为生命科学研究领域的新兴前沿，其中非编码RNA代谢与功能研究是一个核心的基础生物学问题。在前期“基因信息传递过程中非编码RNA的调控作用机制”项目支持取得的基础上，本集成项目将聚焦我国科学家参与发现的新型微小RNA、长非编码RNA和环形RNA等几类非编码RNA，以重要模式生物为研究载体，整合分子、细胞、计算和遗传的技术和方法，研究它们的加工、成熟、结构、降解和功能机制等非编码RNA的基础生物学问题，揭示其加工代谢与功能调控的分子机制。研究结果将揭示非编码RNA代谢的基本规律，发现不同类型的非编码RNA分子家族的新功能元件及由其引发的生命活动规律。

近年来研究发现非编码RNA（ncRNA）参与了多种生命活动的调控，然而其生成、加工、结构与功能机制还缺乏深入研究。近年来，随着高通量测序技术和现代生物学研究手段的飞速发展，多来源、多类型的ncRNA在人类和动植物中被发现，并调控包括免疫应答、癌症、生殖发育等多种生理和病理过程。因此，对ncRNA代谢与功能的研究具有重要意义。本集成项目整合了国内研究领域的多位优秀科学家团队，聚焦人、动物、重要模式生物和植物体内 ncRNA的加工、成熟、结构、降解和功能机制研究，取得了以下一系列重要成果：1）在环形RNA生成、代谢与翻译调控方面:①揭示了内含子来源环形RNA降解与功能耦联新机制，②解析了环形RNA翻译的调控网络机制，③发现了驱动环形RNA翻译的短序列元件，④发现了一类线粒体编码的环形RNA，⑤发现了外显子环形RNA出核转运新机制；2）在环形RNA功能与应用方面:①建立了基于CRISPR-Cas13系统的功能环形RNA筛选新体系，并发现具有重要功能的环形RNA，②开发了体外合成环形RNA的免疫原性与作为RNA适配体的潜在应用前景，③揭示了保守环形RNA circboule在生殖过程中的重要功能；3）在植物与秀丽线虫siRNA研究中:①发现了植物体内源siRNA生成调控新机制，②解析了susi-5参与秀丽线虫risiRNA生成的机制；4）在对eRNA的研究中发现了一种原钙蛋白基因簇中增强子的反义eRNA，并解析其参与基因组折叠的机制；5）在RNA可变剪接与功能研究中:①揭示了RNA二级结构介导Dscam可变剪接的分子机制，②发现了Dscam可变剪接在神经系统中的功能机制。本项目的开展发现了多种不同类型ncRNA代谢的基本规律与新的功能机制，进一步加深了对基因组中不同类型ncRNA在生命调控中的生理功能与作用机制的认知，同时为进一步利用ncRNA提供理论基础。