The Construction Of MiRNA Expression Library For Human

Objective: It was reported there were not only mRNAs in cell, but also massively nocoding RNAs. Although they do not code any proteins, the nocoding RNAs participate in the regulation of gene expression. Recently it was confirmed that miRNA(microRNA) is one kind of important nocoding RNAs. miRNA extensively take part in the growth and development of organism and the formation of disease. However the function of these molecules still remains to be determined. In order to investigated their function, we constructed a library of vectors expressing miRNAs.Methods: The target DNA fragments were amplified from HepG2 cell genome, with the primers designed according to Sanger miRBase, and then cloned into plasmid vector. The ability of some miRNA vectors to express miRNA were detected with Northern blot and dual luciferase assays.Result: A library of vectors expressing miRNAs ,including 207 kinds of human miRNAs were constructed. Pre-miRNA and mature miRNA could be expressed in HEK293 from these vectors, such as pmiR-16,pmiR-24 and pmiR-125a. pmiR-16 and pmiR-483 could be expressed in HepG2 cell and inhibit the expression of luciferase tagged with sequence complementary to corresponding miRNA.Conclusion: A large library of vectors expression miRNA was successfully constructed. The functional miRNA can be expressed, from these miRNA expression vectors. Therefore they can be used to screen for functional miRNA.