Lsp2-AS在果蝇Toll通路免疫响应中的调控作用研究

LncRNA (long noncoding RNA) plays an important regulatory role in many life activities, but the study of lncRNA involvement in immune regulation of Drosophila is still not reported. Previously, we have confirmed that lncRNA (Lsp2-AS) can regulate the expression of antibacterial peptides of DrosophilaToll singal pathway in vitro. It is speculated that Lsp2-AS may be involved in the regulation of Toll signal pathway in Drosophila, but its mechanism still needs further study. Therefore, this project will primarily address the localization analysis of Lsp2-AS in Drosophila cells and provide direction for the follow-up experiments; combine CHIRP technology, RIP technology, high-throughput sequencing technology and related bioinformatics analysis to find the target gene and/or target protein of Lsp2-AS; and based on this, further validate that Lsp2-AS regulated target gene or target protein involved in Drosophila Toll immune response in vitro and in vivo experiments; finally reveale that the regulatory roles of Lsp2-AS involved in innate immune response of Drosophila Toll signal pathway. This study has important scientific implications for elucidating the mechanism of lncRNAs regulating innate immune responses in Drosophila melanogaster, as well as understanding the mechanism of innate immune homeostasis in mammals.

lncRNA（long noncoding RNA）在多种生命活动过程中起着重要的调控作用，但lncRNA参与果蝇免疫调控的研究至今仍无报道。前期我们在体外证实了lncRNA（Lsp2-AS）能够调控果蝇Toll通路下游抗菌肽的表达，推测Lsp2-AS可能参与了果蝇Toll通路先天免疫响应的调控，但其机制仍需进一步深入研究。为此，本项目首要解决Lsp2-AS在果蝇细胞内的定位分析，为后续实验的开展提供方向；结合CHIRP技术、RIP技术、高通量测序技术和相关生物信息学分析发现Lsp2-AS作用的靶基因和/或靶蛋白；并在此基础上，进一步对Lsp2-AS调控靶基因或靶蛋白参与果蝇Toll免疫响应进行体内外实验验证，揭示Lsp2-AS在果蝇Toll通路免疫响应中的调控机制。项目的完成对阐明lncRNA调控果蝇先天免疫响应的机制，以及对深入理解哺乳动物先天免疫稳态维持的机制都具有重要的科学意义。

lncRNA（long noncoding RNA）在多种生命活动过程中起着重要的调控作用，但lncRNA参与果蝇免疫调控的研究至今鲜有报道。通过本项目的研究，我们在体外证实了果蝇lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942能够调控果蝇Toll通路下游抗菌肽的表达，并进一步深入研究lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942参与果蝇Toll通路先天免疫响应的调控机制。本项目首要解决lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942在果蝇细胞内的定位分析，为后续实验的开展提供方向；结合CHIRP技术、RIP技术、高通量测序技术和相关生物信息学分析发现lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942作用的靶基因和/或靶蛋白；并在此基础上，进一步对lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942调控靶基因或靶蛋白参与果蝇Toll免疫响应进行体内外实验验证，揭示lncRNA-CR11538、lncRNA-CR46018、lncRNA-CR33942在果蝇Toll通路免疫响应中的调控机制。项目的完成对阐明lncRNA调控果蝇先天免疫响应的机制，以及对深入理解哺乳动物先天免疫稳态维持的机制都具有重要的科学意义。