Inhibitory effect of Ca2+ on in vivo gene transfer by electroporation.
OBJECTIVE
To investigate the specific effects of Ca2+ on transgene expression during electroporation-mediated gene transfer in mice.


METHODS
Skeletal muscle and skin were subjected to in vivo electroporation with a luciferase reporter plasmid, with or without Ca2+ and various other ions.


RESULTS
For in vivo electroporation, the presence of just 10 mmol/L Ca2+ in the DNA solution drastically reduced the resulting transgene expression, to less than 5% of control values. Only Ca2+, not other ions, caused inhibition, and the effect was not tissue specific. More surprisingly, even when Ca2+ ions were delivered by electroporation before or after DNA administration, similar effects were still observed.


CONCLUSIONS
The inhibitory effect of Ca2+ on in vivo gene transfer by electroporation is specific, ie, the inhibitory effect may be related to the cell membrane properties after electroporation and the subsequent resealing event.
School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China.
Yong-gang
YG
Zhao
Hui-li
HL
Lu
Jin-liang
JL
Peng
Yu-hong
YH
Xu
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D018274:Electroporation; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000378:metabolism*
D004247:DNA; D008156:Luciferases; D002118:Calcium