Electroporation of Plasmid DNA to Swine Muscle

Bodles-Brakhop, Angela M.; Draghia-Akli, Ruxandra; Broderick, Kate E.; Khan, Amir S.

doi:10.1007/978-1-61737-982-6_16

Cited by 4 publications

(4 citation statements)

References 18 publications

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“…GLuc is non-native to the mammalian system. This allows for more definitive quantification than other enzymatic biomarkers, such as secreted embryonic alkaline phosphatase, which can have innate levels in vivo [10]. …”

Section: Introductionmentioning

confidence: 99%

Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression

et al. 2017

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BackgroundThe Gaussia princeps luciferase is used as a stand-alone reporter of transgene expression for in vitro and in vivo expression systems due to the rapid and easy monitoring of luciferase activity. We sought to simultaneously quantitate production of other recombinant proteins by transcriptionally linking the Gaussia princeps luciferase gene to other genes of interest through the foot-and-mouth disease virus 2A translational interrupter sequence.ResultsWe produced six plasmids, each encoding a single open reading frame, with the foot-and-mouth disease virus 2A sequence placed either N-terminal or C-terminal to the Gaussia princeps luciferase gene. Two plasmids included novel Gaussia princeps luciferase variants with the position 1 methionine deleted. Placing a foot-and-mouth disease virus 2A translational interrupter sequence on either the N- or C-terminus of the Gaussia princeps luciferase gene did not prevent the secretion or luminescence of resulting chimeric luciferase proteins. We also measured the ability of another polycistronic plasmid vector with a 2A-luciferase sequence placed downstream of the foot-and-mouth disease virus P1 and 3C protease genes to produce of foot-and-mouth disease virus-like particles and luciferase activity from transfected cells. Incorporation of the 2A-luciferase sequence into a transgene encoding foot-and-mouth disease virus structural proteins retained luciferase activity and the ability to form virus-like particles.ConclusionsWe demonstrated a mechanism for the near real-time, sequential, non-destructive quantitative monitoring of transcriptionally-linked recombinant proteins and a valuable method for monitoring transgene expression in recombinant vaccine constructs.

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Section: Introductionmentioning

confidence: 99%

Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression

et al. 2017

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“…This makes such local delivery methods clinically irrelevant for diseases affecting large, profound and/or multiple muscles . Furthermore, issues of scaling‐up from mouse to bigger animals have been reported . (iii) Muscle electroporation generally consists of IM injections, followed by the application of a pulsatile electric field delivered by electrodes positioned on one or both sides of the muscle area to be treated.…”

Section: Introductionmentioning

confidence: 99%

“…(iii) Muscle electroporation generally consists of IM injections, followed by the application of a pulsatile electric field delivered by electrodes positioned on one or both sides of the muscle area to be treated. This allows a 10‐ to 100‐fold increased transfection compared to IM injection in small and in large animals, as well as in humans. Nevertheless, only mm 3 to cm 3 volumes of muscle can be efficiently treated with single procedures.…”

Section: Introductionmentioning

confidence: 99%

Gene transfer to skeletal muscle using hydrodynamic limb vein injection: current applications, hurdles and possible optimizations

Guen

Gall

Midoux

et al. 2020

The Journal of Gene Medicine

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Hydrodynamic limb vein injection is an in vivo locoregional gene delivery method. It consists of administrating a large volume of solution containing nucleic acid constructs in a limb with both blood inflow and outflow temporarily blocked using a tourniquet. The fast, high pressure delivery allows the musculature of the whole limb to be reached. The skeletal muscle is a tissue of choice for a variety of gene transfer applications, including gene therapy for Duchenne muscular dystrophy or other myopathies, as well as for the production of antibodies or other proteins with broad therapeutic effects. Hydrodynamic limb vein delivery has been evaluated with success in a large range of animal models. It has also proven to be safe and welltolerated in muscular dystrophy patients, thus supporting its translation to the clinic.However, some possible limitations may occur at different steps of the delivery process. Here, we have highlighted the interests, bottlenecks and potential improvements that could further optimize non-viral gene transfer following hydrodynamic limb vein injection. K E Y W O R D Sgene transfer, hydrodynamic delivery, locoregional, non-viral gene delivery, optimizations, skeletal muscle

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“…This results in many cases in the improvement of DNA vaccine efficacies [3][4][5][6]. In particular, electroporation has been demonstrated as a powerful technique also in large animals, including pigs [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Targeted Collection of Plasmid DNA in Large and Growing Animal Muscles 6 Weeks after DNA Vaccination with and without Electroporation

Dory

Moigne

Cariolet

et al. 2015

Journal of Immunology Research

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DNA vaccination has been developed in the last two decades in human and animal species as a promising alternative to conventional vaccination. It consists in the injection, in the muscle, for example, of plasmid DNA encoding the vaccinating polypeptide. Electroporation which forces the entrance of the plasmid DNA in cells at the injection point has been described as a powerful and promising strategy to enhance DNA vaccine efficacy. Due to the fact that the vaccine is composed of DNA, close attention on the fate of the plasmid DNA upon vaccination has to be taken into account, especially at the injection point. To perform such studies, the muscle injection point has to be precisely recovered and collected several weeks after injection. This is even more difficult for large and growing animals. A technique has been developed to localize precisely and collect efficiently the muscle injection points in growing piglets 6 weeks after DNA vaccination accompanied or not by electroporation. Electroporation did not significantly increase the level of remaining plasmids compared to nonelectroporated piglets, and, in all the cases, the levels were below the limit recommended by the FDA to research integration events of plasmid DNA into the host DNA.

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Electroporation of Plasmid DNA to Swine Muscle

Cited by 4 publications

References 18 publications

Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression

Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression

Gene transfer to skeletal muscle using hydrodynamic limb vein injection: current applications, hurdles and possible optimizations

Targeted Collection of Plasmid DNA in Large and Growing Animal Muscles 6 Weeks after DNA Vaccination with and without Electroporation

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