Selective Knockdown of Angiotensin-converting Enzyme In Rat Vascular Smooth Muscle Cells by RNA Interference
Zhou Hua
Zhang Cuifang
Cheng Ruirui
Gao Fen
Yang Zhiming
Abstract：Objective To construct adenovirus vector carrying the shRNA of rat angiotensin-converting enzyme (ACE).To selectively knock down the expression of angiotensin-converting enzyme (ACE) in rat vascular smooth muscle cells by RNA interference.Methods The rat ACE-shRNA segments was obtained from plasmid p-ACE-shRNA which was constructed at an earlier date by RT-PCR and then was cloned into the shuttle plasmid pDC316 to form the pDC316-EGFP-ACE-shRNA vector.The pDC316-EGFP-ACE-shRNA plasmid was cotransfected with genomic plasmid pBHGlox-E1,3Cre into 293 cells to package the recombinant adenovirus.The recombinant adenovirus was transfected into the primary cultured rat vascular smooth muscle cells.ACE mRNA expression were analyzed from rat vascular smooth muscle cells before transfection and 24,48,72 hours after transfection by real-time fluorescence quantitative PCR.Results Recombinant adenoviral vector Ad-EGFP-ACE-shRNA was constructed successfully,which was confirmed by restriction enzyme digestion,PCR and GFP expression.Expression of ACE mRNA in rat vascular smooth muscle cells were significantly reduced 48 hours after Ad-EGFP-ACE-shRNA transfection,and was nearly undetectable after 72 hours.Conclusion The recombinant adenoviral vector carrying ACE-shRNA is successfully constructed.RNA interference can knockdown ACE expression in cultured rat vascular smooth muscle cells,which maybe provide.a new gene therapy idea for cardiovascular diseases.
Keywords：RNA interferencevascular smooth muscle cellsangiotensin-converting enzyme
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 2821-2826 )