lncRNA-BMNCR/miR-145/FSCN1途径调控奶牛金葡菌型乳腺炎自体保护的分子机制

研究奶牛乳腺炎发生发展过程中自体保护的分子机制对于乳腺炎防控具有重要意义。申请人前期鉴定到一个在奶牛金葡菌型乳腺炎中上调表达的长链非编码RNA-BMNCR，生物信息学分析显示BMNCR上存在两个miR-145结合位点。但BMNCR在奶牛乳腺炎发生发展过程中行使什么功能？能否通过竞争性结合miR-145，调控miR-145靶基因FSCN1，参与机体免疫应答尚不清楚。因此，本项目拟通过RNAi、western blot和CRISPR/Cas9等试验，阐明BMNCR和miR-145在免疫应答、自体保护中的功能；利用双荧光素酶报告系统、RIP、RNA Pull-down等技术，揭示BMNCR/miR-145/FSCN1间的调控关系，解析BMNCR/miR-145/FSCN1信号转导网络在奶牛乳腺炎发生发展过程中，参与自体保护、诱导免疫应答的分子机制，为后续防控奶牛乳腺炎、开发靶向药物提供重要参考。

Understanding the molecular mechanism of cow mastitis will be helpful for prevention and treatment of mastitis. Our previous study has identified a novel long non-coding BMNCR which differentially expressed in Streptococcus aureus-induced mastitis. Bioinformatics analyses revealed that there are two miR-145 binding sites on BMNCR sequence. However, it is still unclear that what is the exact function of BMNCR, and if BMNCR can act as a competitive endogenous RNA to regulate FSCN1 expression by sponging miR-145 in cow mastitis. Therefore, in the present study, we will reveal the function of BMNCR and miR-145 in immune response and autoprotection through RNAi, western blot and CRISPR/Cas9 tests. Furthermore, by conducting dual luciferase reporter system, RIP and RNA Pull down experiments, we will clarify the regulatory relationship between BMNCR/miR-145/FSCN1 and demonstrate the molecular mechanism of the BMNCR/miR-145/FSCN1 signal transduction network regulating cow mastitis and immune response. These efforts will provide an important reference for further prevention and treatment of mastitis and the development of targeted drugs in dairy cows.