Abstract
We generated transient transgenic zebrafish by applying electrical pulses subsequent to injection of DNA into muscle tissue of 3–6-month old adult zebrafish. Electroporation parameters, such as number of pulses, voltage, and amount of plasmid DNA, were optimized and found that 6 pulses of 40 V/cm at 15 μg/fish increased the luciferase expression by 10-fold compared with those in controls. By measuring the expression of luciferase, in vivo by electroporation in adult zebrafish and in vitro using fish cell line (Xiphophorus xiphidium A2 cells), the strength of three promoters (CMV, human EF-1α, and Xenopus EF-1α) was compared. Subsequent to electroporation after injecting DNA in the mid region of zebrafish, expression of green fluorescent protein was found far away from the site of injection in the head and the tail sections. Thus, electroporation in adult zebrafish provides a rapid way of testing the behavior of gene sequences in the whole organism.
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References
1. Bigey, P., Bureau, M.F., and Scherman, D. (2002) In vivo plasmid DNA electrotransfer. Curr. Opin. Biotechnol. 13, 443–447.
2. Herweijer, H. and Wolff, J.A. (2003) Progress and prospects: naked DNA gene transfer and therapy. Gene Ther. 10, 453–458.
3. Fattori, E., La Monica, N., Ciliberto, G., and Toniatti, C. (2002) Electro-gene-transfer: a new approach for muscle gene delivery. Somat. Cell Mol. Genet. 27, 75–83.
4. Neumann, C.J. (2002) Vertebrate development: a view from the zebrafish. Semin. Cell Dev. Biol. 13, 469.
5. Powers, D.A., Hereford, L., Cole, T., et al. (1992) Electroporation: a method for transferring genes into the gametes of zebrafish (Brachydanio rerio), channel catfish (Ictalurus punctatus), and common carp (Cyprinus carpio). Mol. Mar. Biol. Biotechnol. 1, 301–308.
6. Sudha, P.M., Low, S., Kwang, J., and Gong, Z. (2001) Multiple tissue transformation in adult zebrafish by gene gun bombardment and muscular injection of naked DNA. Mar. Biotechnol. (NY) 3, 119–125.
7. Tan, J.H. and Chan, W.K. (1997) Efficient gene transfer into zebrafish skeletal muscle by intramuscular injection of plasmid DNA. Mol. Mar. Biol. Biotechnol. 6, 98–109.
8. Tawk, M., Tuil, D., Torrente, Y., Vriz, S., and Paulin, D. (2002) High-efficiency gene transfer into adult fish: a new tool to study fin regeneration. Genesis. 32, 27–31.
9. Sambrook, J., Fritsch, E.F., and Maniatis, T. (eds.) (1989) Molecular cloning. A laboratory manual, part II. Cold Spring Harbor Laboratory Press, New York, NY.
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© 2008 Humana Press
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Rao, N.M., Rambabu, K.M., Rao, S.H. (2008). Electroporation of Adult Zebrafish. In: Li, S. (eds) Electroporation Protocols. Methods in Molecular Biology™, vol 423. Humana Press. https://doi.org/10.1007/978-1-59745-194-9_21
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DOI: https://doi.org/10.1007/978-1-59745-194-9_21
Publisher Name: Humana Press
Print ISBN: 978-1-58829-877-5
Online ISBN: 978-1-59745-194-9
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