Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart
Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart
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DOI：
10.1016/j.bbrc.2005.08.052
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发表时间：
2005-10-14
影响因子：
3.1
通讯作者：
Yoshikawa, T
中科院分区：
文献类型：
--
作者：
Tsunoda, S;Mazda, O;Yoshikawa, T
使用第三代微泡 BR14 通过声孔作用将裸质粒 DNA (pDNA) 和短干扰 RNA (siRNA) 双链体转导到成年小鼠心脏中。将携带荧光素酶、β-半乳糖苷酶 (β-gal) 或增强型绿色荧光蛋白 (EGFP) 报告基因的质粒 DNA 与 BR14 混合，并经皮注射到 C57BL/6 小鼠的左心室 (LV) 腔中，同时暴露于 1 MHz 的经胸超声 60 秒。输出强度为 2.0 W/cm(2) 和脉冲占空比为 50% 的声孔处理导致心脏中荧光素酶表达最高。组织学检查显示左室心内膜下心肌中 P-gal 和 EGFP 报告基因显着表达。在 EGFP 转基因小鼠中，脑室内共注射 siRNA-GFP 和 BR 14 并同时进行超声波照射，导致冠状动脉中 EGFP 表达大幅减少。本方法可适用于成年小鼠心脏的体内功能获得和功能丧失基因工程。 (c) 2005 Elsevier Inc. 保留所有权利。
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 1 MHz for 60 s. Sonoporation at an output intensity of 2.0 W/cm(2) and a 50% pulse duty ratio resulted in the highest luciferase expression in the heart. Histological examinations revealed significant expression of the P-gal and EGFP reporters in the subendocardial myocardiurn of LV. Intraventricular co-injection of siRNA-GFP and BR 14 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. (c) 2005 Elsevier Inc. All rights reserved.