罗氏沼虾性腺发育相关microRNA的鉴定及其调控机制的研究

脊椎动物性腺分化发育过程中microRNA(miRNA)对其靶基因的转录后调控研究较为丰富，但在虾蟹类中报道较少。罗氏沼虾性腺分化发育直接影响其苗种产量和质量，而miRNA对罗氏沼虾性腺分化发育调控的研究仍未开展。本课题以罗氏沼虾性腺差异表达的miRNA为主要研究对象，探究miRNA在罗氏沼虾性腺分化发育过程中的表达规律及调控功能。通过高通量测序，比对分析性腺分化后新表达的及性腺不同发育期特异表达的miRNA和mRNA；并利用生物信息学、定量PCR、原位杂交和RNA干扰等方法预测和初步验证miRNA与其靶基因的功能；然后比较性腺不同发育时期miRNA的表达规律；最后分别通过体外和体内注射miRNA agomir和miRNA antagomir研究miRNA对靶基因的调控机制，以期在罗氏沼虾中建立以miRNA为重要节点的性腺分化发育的调控通路。

During gonadal differentiation and development (D&D) in vertebrate, many reports suggested that the microRNA (miRNA) plays an essential role in the regulation of target genes through the mechanism of post-transcription. However, the correspondent information is rarely characterized in shrimps and crabs. Although gonadal D&D affect yield and quality of Macrobrachium rosenbergii larva directly, study of miRNA regulation in this field has not been carried out until now. In this project, we will mainly focus on the differently expressed miRNA in the gonad to explore its expression profile and regulation function during gonadal D&D in M. rosenbergii. Using next generation high-throughput sequencing technique, we will analyze the newly expressed and unique miRNA and mRNA after gonadal differentiation and different stages of gonadal development, respectively; And prediction and preliminary verification of function of miRNA and its target will be performed by bioinformatics, qPCR, in situ hybridization and RNAi; Then compare the expression patterns of miRNA at different stages of gonadal development; Lastly, in vitro and in vivo injections by miRNA agomir and miRNA antagomir will be performed to research on regulation of miRNA to its target gene, to facilitate a gene regulation pathway of gonadal D&D constructed by miRNA as an important node in M. rosenbergii.