Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10768267 | Biochemical and Biophysical Research Communications | 2005 | 10 Pages |
Abstract
Naked plasmid DNA (pDNA) and short interfering RNA (siRNA) duplexes were transduced into adult murine heart by means of sonoporation using the third-generation microbubble, BR14. Plasmid DNAs carrying luciferase, β-galactosidase (β-gal), or enhanced green fluorescent protein (EGFP) reporter genes were mixed with BR14 and injected percutaneously into the left ventricular (LV) cavity of C57BL/6 mice while exposed to transthoracic ultrasound at 1 MHz for 60 s. Sonoporation at an output intensity of 2.0 W/cm2 and a 50% pulse duty ratio resulted in the highest luciferase expression in the heart. Histological examinations revealed significant expression of the β-gal and EGFP reporters in the subendocardial myocardium of LV. Intraventricular co-injection of siRNA-GFP and BR14 with concomitant ultrasonic exposure resulted in substantial reduction in EGFP expression in the coronary artery in EGFP transgenic mice. The present method may be applicable to gain-of-function and loss-of-function genetic engineering in vivo of adult murine heart.
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Authors
Sei Tsunoda, Osam Mazda, Yohei Oda, Yasunori Iida, Satoshi Akabame, Tsunao Kishida, Masaharu Shin-Ya, Hidetsugu Asada, Satoshi Gojo, Jiro Imanishi, Hiroaki Matsubara, Toshikazu Yoshikawa,