Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection
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 ver...
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ftoslouniv:oai:www.duo.uio.no:10852/46308 2023-05-15T15:28:27+02:00 Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection Schiøtz, Berit L Rosado, Esther G Baekkevold, Espen S Lukacs, Morten Mjaaland, Siri Sindre, Hilde Grimholt, Unni Gjøen, Tor 2011 http://hdl.handle.net/10852/46308 http://urn.nb.no/URN:NBN:no-50565 https://doi.org/10.1186/1756-0500-4-136 eng eng http://urn.nb.no/URN:NBN:no-50565 BMC Research Notes. 2011 May 06;4(1):136 http://hdl.handle.net/10852/46308 http://dx.doi.org/10.1186/1756-0500-4-136 URN:NBN:no-50565 Fulltext https://www.duo.uio.no/bitstream/handle/10852/46308/1/13104_2010_Article_891.pdf Gjøen et al; licensee BioMed Central Ltd. Attribution 2.0 Generic http://creativecommons.org/licenses/by/2.0/ CC-BY Journal article Tidsskriftartikkel Peer reviewed PublishedVersion 2011 ftoslouniv https://doi.org/10.1186/1756-0500-4-136 2020-06-21T08:48:49Z 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. Article in Journal/Newspaper Atlantic salmon Universitet i Oslo: Digitale utgivelser ved UiO (DUO) BMC Research Notes 4 1 |
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Universitet i Oslo: Digitale utgivelser ved UiO (DUO) |
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ftoslouniv |
language |
English |
description |
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. |
format |
Article in Journal/Newspaper |
author |
Schiøtz, Berit L Rosado, Esther G Baekkevold, Espen S Lukacs, Morten Mjaaland, Siri Sindre, Hilde Grimholt, Unni Gjøen, Tor |
spellingShingle |
Schiøtz, Berit L Rosado, Esther G Baekkevold, Espen S Lukacs, Morten Mjaaland, Siri Sindre, Hilde Grimholt, Unni Gjøen, Tor Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
author_facet |
Schiøtz, Berit L Rosado, Esther G Baekkevold, Espen S Lukacs, Morten Mjaaland, Siri Sindre, Hilde Grimholt, Unni Gjøen, Tor |
author_sort |
Schiøtz, Berit L |
title |
Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
title_short |
Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
title_full |
Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
title_fullStr |
Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
title_full_unstemmed |
Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection |
title_sort |
enhanced transfection of cell lines from atlantic salmon through nucoleofection and antibiotic selection |
publishDate |
2011 |
url |
http://hdl.handle.net/10852/46308 http://urn.nb.no/URN:NBN:no-50565 https://doi.org/10.1186/1756-0500-4-136 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_relation |
http://urn.nb.no/URN:NBN:no-50565 BMC Research Notes. 2011 May 06;4(1):136 http://hdl.handle.net/10852/46308 http://dx.doi.org/10.1186/1756-0500-4-136 URN:NBN:no-50565 Fulltext https://www.duo.uio.no/bitstream/handle/10852/46308/1/13104_2010_Article_891.pdf |
op_rights |
Gjøen et al; licensee BioMed Central Ltd. Attribution 2.0 Generic http://creativecommons.org/licenses/by/2.0/ |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1186/1756-0500-4-136 |
container_title |
BMC Research Notes |
container_volume |
4 |
container_issue |
1 |
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1766358814805721088 |