Jan, 2020
Introduction of a plasmid and a protein into bovine and swine cells by water-in-oil droplet electroporation
Journal of Veterinary Medical Science
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- Volume
- 82
- Number
- 1
- First page
- 14
- Last page
- 22
- Language
- English
- Publishing type
- Research paper (scientific journal)
- DOI
- 10.1292/jvms.19-0475
- Publisher
- Japanese Society of Veterinary Science
Instrument cost is a major problem for the transduction of DNA fragments and proteins into cells. Water-in-oil droplet electroporation (droplet-EP) was recently invented as a low-cost and effective method for the transfection of plasmids into cultured human cells. We here applied droplet-EP to livestock animal cells. Although it is difficult to transfect plasmids into bovine fibroblasts using conventional lipofection methods, droplet-EP enabled us to introduce an enhanced green fluorescent protein (EGFP)-expressing plasmid into bovine earlobe fibroblasts. The optimal transfection condition was 3.0 kV, which allowed 19.1% of the cells to be transfected. For swine earlobe fibroblasts, the maximum transfection efficacy was 14.0% at 4.0 kV. After transfection with droplet-EP, 69.1% of bovine and 76.5% of swine cells were viable. Furthermore, droplet-EP successfully transduced Escherichia coli recombinant EGFP into frozen-thawed bovine sperm at 1.5 kV. Flow cytometry analysis revealed that 71.5% of spermatozoa exhibited green fluorescence after transfection. Overall, droplet-EP is suitable for the transfection of plasmids and proteins into cultured livestock animal cells.
- Link information
- ID information
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- DOI : 10.1292/jvms.19-0475
- Pubmed ID : 31776296
- Pubmed Central ID : PMC6983666