PUBLICATION
Efficient transfection of primary zebrafish fibroblasts by nucleofection
- Authors
- Badakov, R., and Jazwinska, A.
- ID
- ZDB-PUB-081114-16
- Date
- 2006
- Source
- Cytotechnology 51(2): 105-110 (Journal)
- Registered Authors
- Jazwinska, Anna
- Keywords
- Fibroblasts, Primary cells, Transfection, Zebrafish
- MeSH Terms
- none
- PubMed
- 19002901 Full text @ Cytotechnology
Citation
Badakov, R., and Jazwinska, A. (2006) Efficient transfection of primary zebrafish fibroblasts by nucleofection. Cytotechnology. 51(2):105-110.
Abstract
Although various gene delivery techniques are available, their application in zebrafish cell cultures has not been extensively studied. Here, we report that nucleofection of zebrafish primary embryonic fibroblasts results in higher transfection efficiency in comparison to other non-viral gene delivery methods. The transfection was performed using green fluorescent protein (GFP) gene constructs of a different size. Greatest DNA uptake was obtained with 4.9-kb plasmid, resulting in 43% GFP positive cells. Nucleofection with 7.4-kb pH2B-GFP plasmid followed by geneticin (G418) selection was successfully used to establish a cell line expressing nuclear histone 2B-GFP fusion protein. Efficient transfection of zebrafish fibroblasts by nucleofection offers a non-viral technique of plasmid delivery and can be used to overexpress genes of interest in these cells.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping