Application of increased temperature from an exogenous source to enhance gene electrotransfer

The presence of increased temperature for gene electrotransfer has largely been considered negative. Many reports have published on the lack of heat from electrotransfer conditions to demonstrate that their effects are from the electrical pulses and not from a rise in temperature. Our hypothesis was...

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Veröffentlicht in:Bioelectrochemistry (Amsterdam, Netherlands) Netherlands), 2015-06, Vol.103, p.120-123
Hauptverfasser: Donate, Amy, Burcus, Niculina, Schoenbach, Karl, Heller, Richard
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Heller, Richard
description The presence of increased temperature for gene electrotransfer has largely been considered negative. Many reports have published on the lack of heat from electrotransfer conditions to demonstrate that their effects are from the electrical pulses and not from a rise in temperature. Our hypothesis was to use low levels of maintained heat from an exogenous source to aid in gene electrotransfer. The goal was to increase gene expression and/or reduce electric field. In our study we evaluated high and low electric field conditions from 90V to 45V which had been preheated to 40°C, 43°C, or 45°C. Control groups of non-heated as well as DNA only were included for comparison in all experiments. Luciferase gene expression, viability, and percent cell distribution were measured. Our results indicated a 2–4 fold increase in gene expression that is temperature and field dependent. In addition levels of gene expression can be increased without significant decreases in cell death and in the case of high electric fields no additional cell death. Finally, in all conditions percent cell distribution was increased from the application of heat. From these results, we conclude that various methods may be employed depending on the end user's desired goals. Electric field can be reduced 20–30% while maintaining or slightly increasing gene expression and increasing viability or overall gene expression and percent cell distribution can be increased with low viability. •Application of increased temperature from an exogenous source enhances expression from GET.•This method can be used to decrease applied voltage while maintaining total gene expression levels.•Increased viability is seen when using this combination under lower voltage conditions.•Electric fields can be reduced up to 30% while still maintaining high levels of gene expression.
doi_str_mv 10.1016/j.bioelechem.2014.08.007
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Many reports have published on the lack of heat from electrotransfer conditions to demonstrate that their effects are from the electrical pulses and not from a rise in temperature. Our hypothesis was to use low levels of maintained heat from an exogenous source to aid in gene electrotransfer. The goal was to increase gene expression and/or reduce electric field. In our study we evaluated high and low electric field conditions from 90V to 45V which had been preheated to 40°C, 43°C, or 45°C. Control groups of non-heated as well as DNA only were included for comparison in all experiments. Luciferase gene expression, viability, and percent cell distribution were measured. Our results indicated a 2–4 fold increase in gene expression that is temperature and field dependent. In addition levels of gene expression can be increased without significant decreases in cell death and in the case of high electric fields no additional cell death. Finally, in all conditions percent cell distribution was increased from the application of heat. From these results, we conclude that various methods may be employed depending on the end user's desired goals. Electric field can be reduced 20–30% while maintaining or slightly increasing gene expression and increasing viability or overall gene expression and percent cell distribution can be increased with low viability. •Application of increased temperature from an exogenous source enhances expression from GET.•This method can be used to decrease applied voltage while maintaining total gene expression levels.•Increased viability is seen when using this combination under lower voltage conditions.•Electric fields can be reduced up to 30% while still maintaining high levels of gene expression.</description><identifier>ISSN: 1567-5394</identifier><identifier>EISSN: 1878-562X</identifier><identifier>DOI: 10.1016/j.bioelechem.2014.08.007</identifier><identifier>PMID: 25193443</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Cell Line ; Cell Survival ; Electrochemical Techniques - methods ; Electroporation ; Electroporation - methods ; Gene electrotransfer ; Gene Expression ; Gene Transfer Techniques ; Heat ; Keratinocytes ; Luciferases - genetics ; Temperature</subject><ispartof>Bioelectrochemistry (Amsterdam, Netherlands), 2015-06, Vol.103, p.120-123</ispartof><rights>2014 Elsevier B.V.</rights><rights>Copyright © 2014 Elsevier B.V. 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subjects Cell Line
Cell Survival
Electrochemical Techniques - methods
Electroporation
Electroporation - methods
Gene electrotransfer
Gene Expression
Gene Transfer Techniques
Heat
Keratinocytes
Luciferases - genetics
Temperature
title Application of increased temperature from an exogenous source to enhance gene electrotransfer
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