lincRNAs在秀丽线虫重要生理活动中的功能

Long intergenic RNAs (lincRNAs) are a class of long noncoding RNAs (lncRNAs) that are located between genes and without overlap with sequences of the coding genes. We have just published our systematic evaluation of C. elegans lincRNAs for their roles and functional mechanisms in relatively simple physiological activities with a CRISPR knockout mutant library. Under the support of this grant, we are going to apply the reverse genetics strategy of “KO mutants-phenotypic analysis-mechanism research” to screen for lincRNAs with phenotypes in more complex physiological activities, and then to study the functions and functional mechanisms of the lincRNAs with phenotype at the molecular, cellular, and the whole organismal levels. The vital physiological activities examined will include reproduction, metabolism, aging, stress responses, male traits, etc. We will investigate lincRNAs with specific phenotypes for their cis or trans regulatory mechanisms, as well as the regulatory network consisting of these lincRNAs. Our studies will systematically reveal the functions and functional mechanisms of lincRNAs as a subclass of lncRNAs in complex physiological activities of the model organism C. elegans at the whole genome level.

基因间长链非编码RNA（lincRNA）是一类位于基因之间、序列不与编码基因重叠的长链非编码RNA（lncRNA）。在前期我们已发表的lincRNAs参与秀丽线虫相对简单生理活动的研究基础上，本项目将利用CRISPR敲除lincRNAs的秀丽线虫品系库，采用“敲除突变-表型分析-机理研究”的反向遗传学研究手段，筛选在特定复杂生理活动中有相应表型的lincRNAs，并在此基础上系统、深入地研究秀丽线虫分子细胞及动物整体水平上lincRNAs在相应生理活动中的功能及功能机理。我们拟研究的重要生理活动包括：生殖、代谢、衰老、应激、雄虫性状，等。对于研究过程中发现的具有表型的lincRNAs，我们将深入研究其可能发挥顺式或反式调控作用的机理及参与形成的分子调控网络。本项目将提升在全基因组水平上、对lincRNAs这一特定类型lncRNAs在重要模式生物秀丽线虫复杂生理活动中的功能及功能机理的认知。

项目申请时，我们利用CRISPR基因编辑构建了一个较为全面的秀丽线虫lincRNAs敲除突变体库，并解析了lincRNAs在线虫简单生理活动中的作用（Genome Biology，2019）。本项目旨在探索复杂生理活动中lincRNAs的功能和功能机理。依据项目计划，我们发现了一些直接调控线虫应激的lincRNAs, 揭示了多种和线虫应激、代谢、发育、生殖、雄虫性状相关的lincRNAs及它们参与的分子调控网络，包括AGO介导的siRNA调控信号。此外本项目还初步探索了响应环境因素的lincRNAs、组织特异性lincRNAs的表达情况、以及从线虫到哺乳动物保守的lincRNAs。在本项目的支持下，发表标注基因号的论文3篇（其中SCI论文2篇，1篇in press），并参与了第十一届全国核糖核酸（RNA）学术讨论会。根据我们的研究结果，基因间长链非编码RNA-lincRNAs与编码基因相比，更多地作用为响应和调控分子而非组成型分子，在线虫的多种复杂生理活动中发挥调节作用。