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, b-galactosidase (b-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 b-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.

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

“…Further frequent treatments with siRNA may lead to more prominent therapeutic outcomes. Other nonviral procedures, including sonoporation 67 and synthetic vectors, 54,68 may also allow repetitive treatments for a melanoma patient.…”

Gene therapy progress and prospects: Ultrasound for gene transfer

“…Most in vivo studies have concentrated on organs and tissues that are readily imaged by diagnostic ultrasound machines, including skeletal muscle, heart and kidneys. 35,36,[48][49][50][51][52][53][54][55][56][57][58][59][60][61][62] Table 1 summarizes the results of some recent studies in these tissues using reporter genes, 35,36,[48][49][50][51][52][53][54]58,60,61 and broadly similar data have begun to emerge from studies in the vascular, 4,63,64 neurology 65-67 and oncology 18,37,68 fields, the pancreas 69 and in hematology. 70 On first inspection the relatively modest enhancements in plasmid-mediated transfection after USE in vivo may appear disappointing compared with the results obtained in vitro.…”

Key factors that affect sonoporation efficiency in in vitro settings: The importance of standing wave in sonoporation

“…(Current address: Department of Neurosurgery, Osaka University Medical School, 2-2 Yamadaoka, Suita, Oaska, 565-0871, Japan. Phone: +81-6-6879-3652: FAX: +81-6-6879-3659; e-mail : m-kinoshita@nsurg.med.osaka-u.ac.jp) hyperthermia, such as coagulation of tissues [1][2][3][4][5][6][7][8][9][10][11][12], mechnical tissue destruction [13,14], disruption of tight junctions of the vasculature for enhanced local drug delivery [15][16][17][18][19][20], and sonoporation, a transient enhanced permeability of the cell membrane caused by the combination of ultrasound and microbubbles [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38]. As all of these bio-effects can be created non-invasively in vivo, ultrasound is gathering a great attention as a next generation surgical or drug delivery system.…”