Importance of association between permeabilization and electrophoretic forces for intramuscular DNA electrotransfer

Bureau, M.; Gehl, Julie; Deleuze, Virginie; Mir, Lluis M.; Scherman, Daniel

doi:10.1016/s0304-4165(00)00028-3

Cited by 201 publications

(148 citation statements)

References 26 publications

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“…This is in agreement with the principle of electrotransfer, which claims a relation between cell diameter, voltage intensity, and transduction efficacy. 19,24 Our demonstration raises a major problem for longterm expression of a therapeutic gene in the context of degenerating muscle. A number of human neuromuscular disorders are characterized by various degrees of acute or chronic degeneration-regeneration cycles and our results would suggest that in these disorders it will be important to target both muscle fibers and the satellite cells.…”

Section: Figure 7 Lack Of Persistence Of Transduced Fibers In An Acutmentioning

confidence: 99%

“…[17][18][19][20][21][22][23] This physical approach improves naked DNA transfer efficacy by a two-component process consisting of membrane permeabilization and DNA movement enhanced by electrophoresis. 19,24 The in vivo application of electric pulses across muscle tissue instantaneously and temporarily compromises the barrier function of the cell membrane, allowing foreign genetic material to enter.…”

Section: Introductionmentioning

confidence: 99%

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Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

Vilquin

Kennel²,

Paturneau-Jouas

et al. 2001

Gene Ther

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The electrotransfer of naked DNA has recently been adapted to the transduction of skeletal muscle fibers. We investigated the short-and long-term efficacy of this methodology in wild-type animals and in mouse models of congenital muscular dystrophy (dy/dy, dy 2J /dy 2J ), or Duchenne muscular dystrophy (mdx/mdx). Using a reporter construct, the short-term efficacy of fiber transduction reached 40% and was similar in wild-type, dy/dy and dy 2J /dy 2J animals, indicating that ongoing muscle fibrosis was not a major obstacle to the electrotransfer-mediated gene transfer. Although the complete rejection of transduced fibers was observed within 3 weeks in the absence of immunosuppression, the persistency was prolonged over 10 weeks when transient or continuous immunosuppressive regimens were used. Using

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Section: Figure 7 Lack Of Persistence Of Transduced Fibers In An Acutmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

Vilquin

Kennel²,

Paturneau-Jouas

et al. 2001

Gene Ther

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“…43 The first experiments were performed using two classical square wave generators, with LV of 83 ms, a limitation imposed by the devices used, and a manual switch to deliver the LV(s) after the HV. 43 Then a device for the controlled generation of such combinations of HV and LV pulses, with, moreover, a controlled gap between the HV and LV pulses, was prepared by the Faculty of Electrical Engineering of the University of Ljubljana, based on a previously developed electroporator. 44 With such an equipment, the roles of the electric pulses in DNA electrotransfer could actually be analysed.…”

Section: Roles Of the Electric Pulses In Dna Electrotransfermentioning

confidence: 99%

DNA electrotransfer: its principles and an updated review of its therapeutic applications

2004

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“…[10][11][12][13][14][15][16] A new protocol with high transfection efficacy in muscle and skin has recently been described, consisting of a combination of one high-field (HV, B1000 V cm À1 ) and one (or several) low-field (LV, B100 V cm À1 ) pulse. [17][18][19][20][21][22][23] For simplicity, we called all along the paper 'field' what indeed is the voltage to electrode width ratio, that is, an experimentally controlled parameter. In these studies, the authors suggested two functions of these electric pulses.…”

Section: Introductionmentioning

confidence: 99%

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

et al. 2009

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Electrotransfer (electroporation) is recognized as one of the most promising alternatives to viral vectors for transfection of different tissues in vivo for therapeutic purposes. We evaluated the transfection efficiency of reporter genes (green fluorescent protein and luciferase) in murine subcutaneous tumors using different combinations of high-field (HV) (600-1400 V cm À1 , 100 ms, 8 pulses) and low-field (LV) (80-160 V cm À1 , 50-400 ms, 1-8 pulses) pulses and compared it to protocol using eight identical pulses of 600 V cm À1 and 5 ms duration (electro-gene therapy, EGT). Expression of GFP was determined using a fluorescent microscope and flow cytometry and expression of luciferase by measuring its activity using a luminometer. The EGT protocol yielded the highest expression of both reporter genes. However, a careful optimization of combinations of HV and LV pulses may result in similar transfection as EGT pulses. With the combination protocol, relatively high fields of LV pulses were necessary to obtain comparable transfection to the EGT protocol. Expression of reporter genes was higher in B16 melanoma than in SA-1 fibrosarcoma. Our data support the hypothesis that both electropermeabilization and electrophoresis are involved in electrotransfer of plasmid DNA, but demonstrate that these components have to happen at the same time to obtain significant expression of the target gene in tumors.

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Importance of association between permeabilization and electrophoretic forces for intramuscular DNA electrotransfer

Cited by 201 publications

References 26 publications

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

DNA electrotransfer: its principles and an updated review of its therapeutic applications

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

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