title: Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart.
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.
authors.affiliation: Department of Molecular Cardiology and Vascular Regenerative Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
authors.forename: Sei
authors.initials: S
authors.lastname: Tsunoda
authors.forename: Osam
authors.initials: O
authors.lastname: Mazda
authors.forename: Yohei
authors.initials: Y
authors.lastname: Oda
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Iida
authors.forename: Satoshi
authors.initials: S
authors.lastname: Akabame
authors.forename: Tsunao
authors.initials: T
authors.lastname: Kishida
authors.forename: Masaharu
authors.initials: M
authors.lastname: Shin-Ya
authors.forename: Hidetsugu
authors.initials: H
authors.lastname: Asada
authors.forename: Satoshi
authors.initials: S
authors.lastname: Gojo
authors.forename: Jiro
authors.initials: J
authors.lastname: Imanishi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Matsubara
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Yoshikawa
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics*; D005260:Female; D005466:Fluorocarbons / Q000737:chemistry*; D018014:Gene Transfer Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry*; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics*; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: C471302:BR14 contrast agent; D005466:Fluorocarbons; D010743:Phospholipids; D034741:RNA, Small Interfering; D004247:DNA