Related Experiment Video
Updated: Aug 11, 2026
Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons
Published on: October 6, 2020
Inhibitory effect of Ca2+ on in vivo gene transfer by electroporation
Yong-gang Zhao1, Hui-li Lu, Jin-liang Peng
1School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China.
Aim:
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.
Conclusion:
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.
Insights
Calcium (Ca2+) significantly inhibits gene transfer via electroporation in mice, reducing expression to under 5%. This specific inhibitory effect, observed even when Ca2+ is applied separately, may involve cell membrane changes post-electroporation.
Area of Science:
- Molecular Biology
- Biotechnology
- Genetics
Background:
- Electroporation is a key method for gene transfer.
- Understanding factors affecting gene transfer efficiency is crucial for therapeutic applications.
Purpose of the Study:
- To investigate the impact of calcium ions (Ca2+) on transgene expression during electroporation-mediated gene transfer.
- To determine if Ca2+ specifically affects gene delivery efficiency in vivo.
Main Methods:
- In vivo electroporation was performed on skeletal muscle and skin in mice.
- A luciferase reporter plasmid was used to assess transgene expression.
- Experiments included the addition of Ca2+ and other ions to the DNA solution.
Main Results:
- 10 mmol/L Ca2+ drastically reduced transgene expression to less than 5% of control values.
- Only Ca2+, not other tested ions, inhibited expression.
- The inhibitory effect was observed regardless of tissue type and even when Ca2+ was applied before or after DNA delivery.
Conclusions:
- Calcium ions (Ca2+) specifically inhibit in vivo gene transfer via electroporation.
- The inhibitory mechanism may be linked to alterations in cell membrane properties and the resealing process after electroporation.
