长链非编码RNA对飞蝗几丁质代谢通路的调控机理

项目名称： 长链非编码RNA对飞蝗几丁质代谢通路的调控机理

项目编号： No.31472051

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 动物学

项目作者： 杨美玲

作者单位： 中国科学院动物研究所

项目金额： 84万元

中文摘要： 长链非编码RNA（lncRNA）对生物体生长发育具有广泛而复杂的基因表达调控作用。几丁质代谢酶的精确调控和表达对于维持昆虫正常蜕皮发育是必需的。本课题组前期研究结果发现非编码RNA miR-71和miR-263b对飞蝗几丁质代谢通路中的关键酶起调控作用，并且lncRNA在几丁质代谢过程中呈现关联性表达变化。在此基础上本项目立足于研究lncRNA对几丁质代谢通路的调控机理，旨在探索几丁质代谢过程中lncRNA和miRNA之间相互作用网络关系，拟开展1）采用体外荧光素酶，RNA pulldown和Rip等技术，验证lncRNA对几丁质代谢酶的调控作用；2）通过体内RNAi/rescue以及共定位方法，阐明lncRNA对几丁质代谢以及蜕皮发育的影响；3）探索lncRNA介导的miRNA对几丁质代谢的网络调控机理。研究结果对于阐释昆虫几丁质代谢通路调控机理以及害虫的有效控制具有重要科学意义。

中文关键词： 长链非编码RNA；几丁质代谢；飞蝗；蜕皮；调控

英文摘要： Long non-coding RNA (lncRNA), as a regulator, plays an important role in growth and development of organisms. Chitin metabolic enzymes are the critical genes involved in the chitin biosynthesis and degradation pathway of insects. The expression of these enzymes is precisely regulated in the normal molting and development of insects. Our previous results showed that miR-71 and miR-263b regulated the chitin metablic pathway by directly targeting two key genes in the migratory locust, and the lncRNAs exprssion was corelated with chitin metabolic enzymes involved in the chitin metabolism process. In this project, we focus on the regulation of lncRNA in the chitin metabolic pathway, providing insights of the molecular interactions between lncRNAs and miRNAs associated with the chitin metabolism. Furthermore, we predicted the potential lncRNAs that target the adjacent chitin metabolic enzymes and analyzed the expression correlation between lncRNAs and chitin metabolic enzymes in chitin metabolism. This project is intended 1) to determine the interactions between lncRNAs and chitin metabolic enzymes by luciferase experiments in vitro, RNA pulldown and RNA-binding protein immunoprecipitation experiments; 2) to prove whether lncRNAs regulate the chitin metabolism and the molting development of metabolic enzymes by RNAi, rescue and in situ hybridization and immunohistochemistry analyses; 3) to verify the role of miRNAs and lncRNAs in the regulation of chitin metabolism by targeting of chitin metabolic enzymes. This study will reveal the novel function and modulation pattern of lncRNAs, providing important insights into the underlying molecular mechanisms of chitin metabolism in locusts.

英文关键词： long non-coding RNA;chitin metabolism;locusta migratoria;molting;regulation