Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart |
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Authors: | Tsunoda Sei Mazda Osam Oda Yohei Iida Yasunori Akabame Satoshi Kishida Tsunao Shin-Ya Masaharu Asada Hidetsugu Gojo Satoshi Imanishi Jiro Matsubara Hiroaki Yoshikawa Toshikazu |
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Affiliation: | Department of Molecular Cardiology and Vascular Regenerative Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan. |
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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, beta-galactosidase (beta-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 1MHz for 60s. Sonoporation at an output intensity of 2.0W/cm(2) and a 50% pulse duty ratio resulted in the highest luciferase expression in the heart. Histological examinations revealed significant expression of the beta-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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Keywords: | Naked plasmid DNA Heart Sonoporation Microbubble Gene transfer RNA interference Ultrasound Gene therapy Molecular therapy |
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