Successful and optimized in vivo gene transfer to rabbit carotid artery mediated by electronic pulse

Matsumoto, Takuya; Komori, Kimihiro; Shoji, Tetsuya; Kuma, Sosei; Kume, Makoto; Yamaoka, Terutoshi; Mori, Emiko; Furuyama, Tadashi; Yonemitsu, Yoshikazu; Sugimachi, Keizō

doi:10.1038/sj.gt.3301502

Cited by 46 publications

(41 citation statements)

References 29 publications

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“…As with adventitial delivery and electroporation, transfer is detected within the endothelial cells and both the intimal and medial smooth muscle layers. Another group delivered DNA with a double balloon catheter and applied the electric field from the adventitial surface of the rabbit carotid with two "T"-shaped electrodes (2.5 cm long ϫ 0.5 cm wide) placed on either side of the vessel segment (82). In both of these cases, gene transfer was dependent on the electric field.…”

Section: Electroporation To Target Smooth Musclementioning

confidence: 99%

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

Dean¹

2005

American Journal of Physiology-Cell Physiology

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The study of muscle physiology has undergone many changes over the past 25 years and has moved from purely physiological studies to those intimately intertwined with molecular and cell biological questions. To ask these questions, it is necessary to be able to transfer genetic reagents to cells both in culture and, ultimately, in living animals. Over the past 10 years, a number of different chemical and physical approaches have been developed to transfect living skeletal, smooth, and cardiac muscle systems with varying success and efficiency. This review provides a survey of these methods and describes some more recent developments in the field of in vivo gene transfer to these various muscle types. Both gene delivery for overexpression of desired gene products and delivery of nucleic acids for downregulation of specific genes and their products are discussed to aid the physiologist, cell biologist, and molecular biologist in their studies on whole animal biology.

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Section: Electroporation To Target Smooth Musclementioning

confidence: 99%

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

Dean¹

2005

American Journal of Physiology-Cell Physiology

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“…Recently, electroporation has been used in the rabbit carotid and has shown similar high-level gene expression and therapeutic efficacy when DNA was delivered intraluminally and the electric field was applied from the adventitial surface. 27 Thus, the use of electroporation for gene transfer to the vasculature is a safe and powerful tool that can be utilized for the study of numerous physiological and cellular processes within the context of the whole animal as well as investigating and exploiting the process of exogenous DNA nuclear import for the development and improvement of nonviral gene therapy vectors.…”

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confidence: 99%

Effect of a DNA nuclear targeting sequence on gene transfer and expression of plasmids in the intact vasculature

2003

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Although the use of nonviral vectors for gene therapy offers distinct advantages including the lack of significant inflammatory and immune responses, the levels of expression in vivo remain much lower than those obtained with their viral counterparts. One reason for such low expression is that unlike many viruses, plasmids have not evolved mechanisms to target to the nucleus of the nondividing cell. In the absence of mitosis, plasmids are imported into the nucleus in a sequence-specific manner, and we have shown in cultured cells by transfection and microinjection experiments that the SV40 enhancer mediates plasmid nuclear import in all cell types tested (Dean et al., 1999, Exp Cell Res 253: 713-722). To test the effect of this import sequence on gene transfer in the intact animal, we have recently developed an electroporation method for DNA delivery to the intact mesenteric vasculature of the rat. Plasmids expressing luciferase or GFP from the CMV immediate-early promoter/enhancer and either containing or lacking the SV40 enhancer downstream of the reporter gene were transferred to the vasculature by electroporation. When transfected into actively dividing populations of smooth muscle or epithelial cells, the plasmids gave similar levels of expression. By contrast, the presence of the SV40 sequence greatly enhanced gene expression of both reporters in the target tissue. At 2 days post-transfer, plasmids with the SV40 sequence gave 10-fold higher levels of luciferase expression, and at 3 days the difference was over 40-fold. The presence of the SV40 sequence did not simply increase the rate of nuclear import and expression, since expression from the SV40-lacking plasmid did not increase beyond that seen at day 2, the time of maximum expression for either plasmid. In situ hybridization experiments confirmed that the increased gene transfer and expression was indeed due to increased nuclear localization of the delivered SV40 sequence-containing plasmid. Based on these findings, the ability to target DNA to the nucleus can increase gene transfer in vivo and inclusion of the SV40 sequence into plasmids will enhance nonviral gene delivery.

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“…However, another type of square-wave electric pulse was applied to muscles (Aihara and Miyazaki, 1998;Mir et al, 1999), tumors (Rols et al, 1998), liver (Suzuki et al, 1998), and some other tissues (André and Mir, 2004), and was found to be more effective for DNA electrotransfer Heller et al, 2000). These pulses usually are of lower voltage but much longer duration (in the range of tens of milliseconds) (Aihara and Miyazaki, 1998;Rols et al, 1998;Mir et al, 1999;Bettan et al, 2000;Matsumoto et al, 2001). It is assumed that this type of pulse mediates DNA transfer into cells by inducing two distinct effects that include cell permeabilization (like the short pulses) and DNA electrophoretic migration during the delivery of the electric field (Klenchin et al, 1991;Sukharev et al, 1992;Neumann et al, 1996;Mir et al, 1999;Golzio et al, 2002).…”

Section: Introduction Ementioning

confidence: 99%

Electrophoretic Component of Electric Pulses Determines the Efficacy of In Vivo DNA Electrotransfer

et al. 2005

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Efficient DNA electrotransfer can be achieved with combinations of short high-voltage (HV) and long lowvoltage (LV) pulses that cover two effects of the pulses, namely, target cell electropermeabilization and DNA electrophoresis within the tissue. Because HV and LV can be delivered with a lag up to 3000 sec between them, we considered that it was possible to analyze separately the respective importance of the two types of effects of the electric fields on DNA electrotransfer efficiency. The tibialis cranialis muscles of C57BL/6 mice were injected with plasmid DNA encoding luciferase or green fluorescent protein and then exposed to various combinations of HV and LV pulses. DNA electrotransfer efficacy was determined by measuring luciferase activity in the treated muscles. We found that for effective DNA electrotransfer into skeletal muscles the HV pulse is prerequisite; however, its number and duration do not significantly affect electrotransfer efficacy. DNA electrotransfer efficacy is dependent mainly on the parameters of the LV pulse(s). We report that different LV number, LV individual duration, and LV strength can be used, provided the total duration and field strength result in convenient electrophoretic transport of DNA toward and/or across a permeabilized membrane.

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Successful and optimized in vivo gene transfer to rabbit carotid artery mediated by electronic pulse

Cited by 46 publications

References 29 publications

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

Effect of a DNA nuclear targeting sequence on gene transfer and expression of plasmids in the intact vasculature

Electrophoretic Component of Electric Pulses Determines the Efficacy of In Vivo DNA Electrotransfer

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