A novel class of small non-coding RNAs

一类新型小非编码 RNA

• 批准号: RGPIN-2019-06502
• 负责人: Provost, Patrick
• 金额: $ 3.06万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=716070
• 关键词: novel; class; small; non; coding

Central to the flow of genetic information (DNA makes RNA makes protein), small RNA species that do not code for proteins, such as microRNAs, have evolved as key regulators of gene expression; this is required to preserve well-balanced protein expression/levels, to maintain normal cellular functions and, for the organisms, to remain fit and healthy. Contrary to current scientific (mis?)conception, that no RNA of fewer than 16 nucleotides (nt) can be biologically relevant, we serendipitously discovered a 12-nt long RNA species that could compete with the microRNA regulation of mRNA translation into proteins in human cells. Analyzing RNA extracted from 12 different primary and cultured cell types, from 5 different species, in the window of RNA sizes comprised between 8 and 30 nt, we identified unusually little RNAs (ulRNAs) of 12 and 13 nt that were often more abundant than 19 to 24-nt microRNAs. Most of the 12-nt RNAs corresponded to a unique sequence, which we have termed dRNA (“d” stands for dozen, douze or duodecim [in Latin]), whereas most 13-nt RNAs consisted of the dRNA sequence with a C at its 5'end (C-dRNA). The dRNA sequence mapped, in the human and mouse genomes, to the 5' end of 5.8S ribosomal RNA, and thus constitute a ribosomal RNA fragment (rRF). The level of dRNA/C-dRNA was much lower in platelets, which are also known for their very low protein output, suggesting a positive association between dRNA/C-dRNA levels and mRNA translation. With the overall objective of elucidating the molecular biology, role, function and importance of ulRNAs, this proposal aims to delineate the origin, biogenesis pathway and effector protein complexes involved in the role and function of dRNA/C-dRNA in human and mouse cells. To confirm our findings and the authenticity of the dRNA/C-dRNA sequences, we will perform additional RNA-Seq analyses and prioritize the validation of an original, sensitive and specific quantitative PCR (qPCR)-based detection method. This qPCR method will also allow us to detect and monitor dRNA/C-dRNA levels under the various experimental settings that will be deployed to characterize this novel class of unusually little RNAs and to elucidate their role, function and importance in human and mouse cells. Challenging our current scientific beliefs, this research program is expected to confirm the existence of a new class of non-coding RNAs and unveil their role and importance in the increasingly complex mechanisms regulating gene expression in eukaryotes. With this proposal, we hope to establish ulRNAs as a new research area and encourage efforts in this field of RNA biology, in order to improve our understanding of life. Our Institution is closely monitoring our progress and is ready to protect our intellectual property, initially through patenting of our qPCR-based, dRNA/C-dRNA detection method and derived applications, as soon as their function is elucidated.

作为遗传信息流动的中心（DNA制造RNA制造蛋白质），不编码蛋白质的小RNA种类（如microRNA）已进化为基因表达的关键调节因子;这是保持蛋白质表达/水平平衡，维持正常细胞功能以及生物体保持健康和健康所必需的。 与目前的科学（错误？）基于一种新的概念，即少于16个核苷酸（nt）的RNA不可能具有生物学相关性，我们偶然发现了一种12-nt长的RNA种类，它可以与人类细胞中mRNA翻译成蛋白质的microRNA调控竞争。 分析从5个不同物种的12种不同的原代和培养细胞类型中提取的RNA，在RNA大小介于8和30 nt之间的窗口中，我们鉴定了12和13 nt的异常小的RNA（ulRNA），其通常比19至24-nt的microRNA更丰富。大多数12-nt RNA对应于一个独特的序列，我们称之为dRNA（“d”代表十几个，douze或duodecim [拉丁语]），而大多数13-nt RNA由5 '端带有C的dRNA序列组成（C-dRNA）。在人类和小鼠基因组中，该dRNA序列定位于5.8S核糖体RNA的5'端，因此构成核糖体RNA片段（rRF）。 在血小板中，dRNA/C-dRNA的水平要低得多，这也是已知的非常低的蛋白质输出，表明dRNA/C-dRNA水平和mRNA翻译之间的正相关。 本研究的总体目标是阐明ulRNA的分子生物学、作用、功能和重要性，旨在描述与人和小鼠细胞中的dRNA/C-dRNA的作用和功能有关的起源、生物发生途径和效应蛋白复合物。 为了证实我们的发现和dRNA/C-dRNA序列的真实性，我们将进行额外的RNA-Seq分析，并优先验证基于原始、灵敏和特异性定量PCR（qPCR）的检测方法。这种qPCR方法还将使我们能够在各种实验环境下检测和监测dRNA/C-dRNA水平，这些实验环境将用于表征这种新型的异常小的RNA，并阐明它们在人类和小鼠细胞中的作用、功能和重要性。 根据我们目前的科学信念，这项研究计划有望证实一类新的非编码RNA的存在，并揭示它们在真核生物中日益复杂的基因表达调控机制中的作用和重要性。通过这个提议，我们希望将ulRNA建立为一个新的研究领域，并鼓励在RNA生物学这一领域的努力，以提高我们对生命的理解。我们的机构正在密切关注我们的进展，并准备好保护我们的知识产权，首先是通过我们基于qPCR的dRNA/C-dRNA检测方法和衍生应用程序的专利，一旦它们的功能得到阐明。