Construction and identification of recombinant adenovirus expressing short hairpin RNA of rat angiotensin-converting enzyme

Objective To construct an adenovirus vector expressing shRNA of rat angiotensin-converting enzyme(ACE) and amplify the adenovirus vector in HEK-293 cells.Methods The rat ACE-shRNA segments was obtained from plasmid pGenesil-1-ACE-shRNA that was constructed at an earlier date by RT-PCR and then was cloned into the shuttle plasmid pDC316 to form the pDC316-ACE-shRNA vector.The pDC316-ACE-shRNA plasmid was cotransfected with genomic plasmid pBHGlox-E1,3Cre into HEK-293 cells to package the recombinant adenovirus.The recombinant adenovirus was transfected into rat vascular endothelia1 cells(ECs),and the green fluorescence protein expression was detected.Results Recombinant adenoviral vector Ad-ACE-shRNA was constructed successfully,which was confirmed by restriction enzyme digestion,PCR and GFP expression.Conclusion The recombinant adenoviral vector carrying ACE-shRNA was successfully constructed,which would become a basis for gene silence in antihypertensive treatment.