降解组测序技术在植物miRNA研究中的应用

目前, 利用芯片技术和miRNA测序可快速、准确地检测到物种中所含有的miRNA。随着越来越多的miRNA被发现, miRNA靶基因的确定已成为研究miRNA生物学功能的关键。传统的miRNA靶基因的寻找主要依赖生物信息学预测、AGO蛋白免疫共沉淀和荧光素酶法等。随着高通量测序技术的持续革新, 出现了一种新的miRNA靶基因的检测方法, 即降解组测序(degradome sequencing)法, 该方法拥有高通量测序技术、生物信息学分析和RACE验证三者的优势, 并已成功应用于拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)和小立碗藓(Physcomitrella patens)等模式植物miRNA靶基因的检测。基于已发表的相关文献和联川生物降解组测序平台, 该文对降解组测序技术应用于植物miRNA靶基因的研究进展及其实验原理进行了综述, 同时对运用该技术可进行的更深入研究进行了讨论。

Use of Degradome Sequencing in Study of Plant MicroRNAs

MicroRNA (miRNA) chip and sequencing have been used to detect miRNAs in species. With the discovery of more miRNAs, miRNA target genes are the key to study miRNA biological function. Finding miRNA target genes mainly relies on computational prediction, AGO protein co-immunoprecipitation and luciferase. Deep sequencing, or degradome sequencing, can be used for high-throughput identification of targets of miRNAs. The method includes deep sequencing, bioinformatic analysis and 5'-rapid amplification of cDNA ends. It has been successfully used for global identification of miRNA-target RNA pairs in Arabidopsis thaliana, Oryza sativa and Physcomitrella patens. This paper reviews research progress in degradome sequencing in plant miRNA target study and discusses trends in further application of degradome sequencing.