Journal of Southern Medical University ›› 2013, Vol. 33 ›› Issue (02): 202-.

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Construction of adenoviral vectors expressing miR-30a and miR-30e

  

  • Online:2013-02-20 Published:2013-02-20

Abstract: Objective To construct adenoviral vectors expressing mature miRNA-30a and miRNA-30e. Methods The target
mmu-miR-30a and mmu-miR-30e genes amplified from mouse genome were digested and linked to the shuttle plasmid
pSES-HUS, which was then transformed into competent AdEaseier cells for recombination. The confirmed recombinant
plasmids were transfected into Hek-293 cells for production of the adenoviruses pAd-mmu-miR-30a and pAd-mmu- miR-30e.
The obtained adenoviruses were used to infect Mefs cells, and the cellular expressions of mmu-miR-30a and mmu- miR-30e
were detected using fluorescence quantitative PCR. Results mmu-miR-30a (357 bp) and mmu-miR-30e (324 bp) containing the
restriction sites were amplified and linked to the shuttle plasmid pSES-HUS, which was successfully recombined with
AdEasy1. After packaging in Hek-293 cells, the adenoviral vectors were obtained, which caused an increase of mmu-miR-30a
expression by 26.46±7.46 folds and mmu-miR-30e expression by 2.76±0.25 folds in transfected Mefs cells. Conclusion We have
successfully constructed the adenoviral vectors expressing the mature miRNAs.