新型隐球酵母基因间来源的小RNA功能验证

Interfering effect of an intergenic-derived sRNA of Cryptococcus neoformans

【目的】验证内源基因间来源的20-25nt的small RNA(sRNA)是否具有干扰功能。【方法】通过建立隐球酵母sRNA测序文库,得到约200个长度在20-25 nt的sRNAs,运用建立的验证新型隐球酵母sRNA功能报告系统,对其进行干扰功能验证。【结果】通过CLC-sRNA报告系统,对选取的sRNA进行了分子水平的功能验证,发现一个具有干扰功能的基因间来源的sRNA。【结论】新型隐球酵母sRNA来源具有多样性,对报告靶基因存在着干扰功能,其干扰的通路应当是经典RNAi通路,同时在无性传递过程中具有稳定性。

[ Objective ] We demonstrated the interfering effect of a new intergenic-derived sRNA of Cryptococcus neoformans. [Methods] We constructed a library of sRNAs from 20 to 25 nt, and used a reporter gene CLC1 that was fused to the testing sRNAs to show the interference effect, i.e. the degradation of the CLC1 would gene rate an albino phenotype of the transformants. [ Results] Through the CLC-sRNA reporter system, we acquired one sRNA, sRNA-1, located to an intergenic region of the genome of C. neoformans, that displayed interfering effect on CLC1. [ Conclusions] In C. neoformans, the origination of sRNA may be diverse. The endogenous sRNA should play interfering function via the canonical RNAi pathway.