Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection

• Published in: BMC Research Notes
• Main Authors: Mjaaland Siri, Lukacs Morten, Baekkevold Espen S, Rosado Esther G, Schiøtz Berit L, Sindre Hilde, Grimholt Unni, Gjøen Tor
• Format: Article in Journal/Newspaper
• Language: English
• Published: BMC 2011
• Subjects: Medicine; R; Biology (General); QH301-705.5; Science (General); Q1-390; Atlantic salmon
• Online Access: https://doi.org/10.1186/1756-0500-4-136; https://doaj.org/article/46fb9a44f4e94342859a8672f12ee9b3

Abstract Background Cell lines from Atlantic salmon kidney have made it possible to culture and study infectious salmon anemia virus (ISAV), an aquatic orthomyxovirus affecting farmed Atlantic salmon. However, transfection of these cells using calcium phosphate precipitation or lipid-based reagents shows very low transfection efficiency. The Amaxa Nucleofector technology™ is an electroporation technique that has been shown to be efficient for gene transfer into primary cells and hard to transfect cell lines. Findings Here we demonstrate, enhanced transfection of the head kidney cell line, TO, from Atlantic salmon using nucleofection and subsequent flow cytometry. Depending on the plasmid promoter, TO cells could be transfected transiently with an efficiency ranging from 11.6% to 90.8% with good viability, using Amaxa's cell line nucleofector solution T and program T-20. A kill curve was performed to investigate the most potent antibiotic for selection of transformed cells, and we found that blasticidin and puromycin were the most efficient for selection of TO cells. Conclusions The results show that nucleofection is an efficient way of gene transfer into Atlantic salmon cells and that stably transfected cells can be selected with blasticidin or puromycin.