Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart.
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.
Tsunoda
Sei
S
Department of Molecular Cardiology and Vascular Regenerative Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
Mazda
Osam
O
Oda
Yohei
Y
Iida
Yasunori
Y
Akabame
Satoshi
S
Kishida
Tsunao
T
Shin-Ya
Masaharu
M
Asada
Hidetsugu
H
Gojo
Satoshi
S
Imanishi
Jiro
J
Matsubara
Hiroaki
H
Yoshikawa
Toshikazu
T
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
C471302:BR14 contrast agent; D005466:Fluorocarbons; D010743:Phospholipids; D034741:RNA, Small Interfering; D004247:DNA