Corin基因在雪兔季节性毛色变化中的功能研究

动物季节性毛色变化是重要的科学问题 。申请者用RNA-Seq和de nove转录组技术获得了雪兔不同毛色变化期间皮肤组织4个差异表达的基因，这些基因的序列和家兔及野兔比较并无导致蛋白质变异的差异；比较变色兔与非变色兔的基因组序列，发现只有Corin基因的启动区发生了很大的变异。Corin基因敲除小鼠毛色变淡，且在毛乳头表达，具备把内分泌或神经系统信号传递到毛囊色素细胞的条件，因此是唯一的候选基因。为了验证雪兔Corin基因的功能，本项目拟首先获得雪兔Corin基因调控区的序列，其次利用CRISPR/Cas9技术将雪兔Corin基因失活，观察其毛色是否变白；第三利用CRISPR/Cas9技术将雪兔Corin基因调控区的变异序列敲入家兔并置换家兔Corin基因相应的区段，观察毛色是否像雪兔一样出现季节性变化。Corin功能的验证对于揭示其调控毛发色素合成及感受环境变化的机理具有重要的理论意义。

Animal seasonal colour changes is important scientific question. Applicants with RNA-Seq and de nove transcriptome, four skin differential expressed genes were obtained between two stages of coat color change, these gene sequences among mountain hare, rabbits and other hares do not lead to differences in protein variations, compared the genome sequences of hare found that Corin gene have a great variations in the promoter region. Corin knockout mouse cause coat color fades, and Corin gene expressed in dermal papilla, so that it have ability for set up relationship between endocrine or nervous-system signals and hair follicle melanocytes, is the only candidate gene. On the basis of previous studies, to identify function of Corin gene in mountain hare, this project intends first get mountain hare Corin gene sequence of variation region, second using CRISPR/Cas9 technology knock out Corin gene of mountain hare, observation its hair color whether variable white; third using CRISPR/Cas9 technology knock mountain hare Corin gene into rabbit genome and replace the different region of rabbit Corin gene, observation hair color whether like mountain hare as appeared seasonal changes. This research has a great theoretical significance for clarifying the regulation of melanogenesis and reception of evironment change.