家蚕长链非编码RNA参与调控脂肪体发育和细胞自噬的机制研究

长链非编码RNA（lncRNAs）在许多生命活动中发挥重要作用，但其在昆虫发育和细胞自噬中的作用机制还不清楚。前期研究通过转录组测序在蜕皮激素诱导后家蚕脂肪体中筛选到两个lncRNAs，lnc560与自噬相关基因的时空表达高度吻合，lnc585干涉表达可显著影响幼虫到蛹发育，推测它们可能参与家蚕脂肪体发育和细胞自噬的调控。为进一步揭示这些lncRNAs的作用机制，本项目将通过体外实验解析lncRNAs与靶基因的互作机制；进而在家蚕细胞和体外培养脂肪体中抑制和激活lncRNAs表达，研究其通过靶基因调控脂肪体发育和细胞自噬的分子机制；并在个体沉默或敲除lncRNAs，探索两个lncRNAs在家蚕变态发育时期的调控关系。本研究将为揭示lncRNAs参与昆虫变态发育和自噬的调控机制提供新的认识，创新和拓展lncRNAs的调控机制内容，同时为家蚕遗传发育调控和鳞翅目害虫防治提供新的候选靶分子。

Long non-coding RNAs (lncRNAs) play important roles in many biological processes, but the regulatory mechanisms of lncRNAs in insect development and autophagy are still unclear. In previous studies, two lncRNAs were screened from the fat body of silkworm after 20-hydroxyecdysone induction by transcriptome sequencing. Lnc560 was highly consistent with the temporal and spatial expression of autophagy related gene, and interference of lnc585 could significantly affect the development of larva to pupa, which suggested that lncRNAs might be involved in the regualtion of fat body development and autophagy in silkworm, Bombyx mori. To further reveal the regulatory mechanisms of these lncRNAs, we will first analyze the interaction mechanism between lncRNA and its target gene in vitro. Then we will inhibit and activate the expression of lncRNAs in silkworm cell line and fat body cultured in vitro to clarify the molecular mechanism of lncRNAs regulating fat body development and autophagy by target genes. Finally we propose to silence or knock out the expression of lncRNAs to explore the regulatory relationship between two lncRNAs during metamorphosis of silkworm. This research will not only provide a new understanding for the regulatory mechanism of lncRNAs involved in metamorphosis and autophagy in insects, but also innovate and expand the regulatory mechanism of lncRNAs. Meanwhile, it will provide new candidate targets for the genetic development regulation of silkworm and the control of Lepidoptera pests.