绵羊miRNA-411a与其靶基因FLT3的相互作用

Interaction Between miRNA-411a and FLT3 in Sheep

miRNAs是一类长约22nt的非编码单链小RNA分子,主要与靶基因的3′UTR结合参与生物体基因的表达和调控过程。已经证实许多miRNA通过对靶标的调控,在支原体肺炎感染过程中起重要作用。为验证miRNA-411a与预测的靶基因FLT3间存在相互作用,采用生物信息学方法分析预测其二级结构,利用荧光定量PCR技术分析miRNA-411a的正常表达和过量表达后靶基因FLT3的相对表达量,双荧光素酶报告基因试验验证两者的相互作用。结果表明,miRNA-411a与其靶基因FLT3能形成典型的二级茎环结构;在过量表达miRNA-411a后,其靶基因FLT3的表达量显著降低。综上所述,miRNA-411a是通过与FLT3的3′UTR结合而发挥作用,确定[FLT3是miRNA-411a的靶基因。

miRNAs are a class of small non-coding single-stranded RNA molecules with a length of about 22 nt. They mainly participate in the expression and regulation of genes in the organism by binding to the 3′UTR of the target gene. Many miRNAs play an important role in the infection of mycoplasma pneumonia by regulating the target. In order to verify this interaction exists between miRNA-411a and its predicted target gene FLT3, bioinformatics analysis was used in this study to predict its secondary structure, and analyze the normal expression and overexpression of miRNA-411a by real-time PCR. The interaction between miRNA-411a and FLT3 were verified by relative expression levels of the target gene FLT3 and dual luciferase reporter gene experiments. The results showed that miRNA-411a and its target gene FLT3 could form a typical secondary stem-loop structure. After over-expression of miRNA-411a, the expression level of its target gene FLT3 was significantly decreased, and the interaction between miRNA-411a and FLT3 were verified. In summary, miRNA-411a functions by binding to the 3′UTR of FLT3, FLT3 is a target gene of miRNA-411a.