Transfection of Kir6. 2 channel genes into native vascular smooth muscle using sonoporation

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The combination of ultrasound and nano-bubbles coated with lipid bilayers(i. e. hybrid nano-bubbles) induces transient membrane permeability, leading to direct delivery of exogenous molecules(such as plasmid, siRNA, anticancer drugs etc.) into cells with minimal invasion. This method is generally termed as to be a sonoporation technique. When Kir6. 2(inwardly-rectifying K^+ channel family 6 subtype 2) genes have transfected into native vascular smooth muscle layers of mouse aorta by use of sonoporation, RT-PCR analysis revealed the expression of Kir6. 2 transcripts in vascular smooth muscle. When Kir6. 2 genes, tagging with Myc-genes, were transfected into native smooth muscle layers using sonoporation, immunohistochemical studies have revealed that Kir6. 2 and Myc proteins were co-expressed in vascular smooth muscle cells. The phenylephrine-induced contraction of mouse aorta was significantly reduced after the treatment of Kir6. 2 gene-sonoporation, hyperpolaring the membrane potentials of vascular smooth muscle.These results suggest that Kir6. 2 genes were functionally expressed in mouse vascular smooth muscles, causing a vascular relaxation due to the activity of Kir6. 2. Thus, we have succeeded to establish the novel gene-transfected method.