Abstract
DNA delivery into fish is important for transient gene expression, (e.g., DNA vaccination). Previous studies have generally focused on intramuscular injection of DNA vaccines into fish. However, this method is obviously impractical and laborious for injecting large numbers of fishes. This study reports oral delivery of a construct expressing the β-galactosidase reporter gene into fish by encapsulating the DNA in chitosan and incorporating it into fish feeds. We found that β-galactosidase expression could be observed in the stomachs, spleens, and gills of fishes fed with flakes containing the chitosan-DNA complex. These results suggest that DNA vaccines and other constructs can be easily and cheaply delivered into fishes orally by use of carriers and incorporation into fish feeds.
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References
E.D. Anderson D.V. Mourich S.C. Fahrenkrug S La Patra J.C. Leong J. Shepherd (1996) ArticleTitleGenetic immunization of rainbow trout (Oncorhynchus mykiss) against infectious necrosis hematopoietic virus Mol Mar Biol Biotechnol 5 114–122
E.D. Anderson J.C. Leong (2000) Development of DNA vaccines for salmonid fish D.B. Lowrie R.G. Whalen (Eds) DNA vaccines: Methods and Protocols Totowa Humana Press N.J.
S.C. Chen D.H. Jones E.F. Fynan G.H. Farrar J.C.S. Clegg H.B. Greenberg J.E. Herrmann (1997) ArticleTitleProtective immunity induced by oral immunization with a rotavirus DNA vaccine encapsulated in microparticles J Virol 72 5757–5761
S. Corbeil G. Kurath S.E. LaPatra (2000) ArticleTitleFish DNA vaccine against infectious hematopoietic necrosis virus: efficacy of various routes of immunization Fish Shellfish Immunol 10 711–723 Occurrence Handle10.1006/fsim.2000.0286
W.W. Daniel (1991) Biostatistics: A Foundation for Analysis in the Health Sciences John Wiley and Sons Hopoken, N.J.
J.H. Eldridge C.J. Hammond J.A. Meulbroek J.K. Staas R.M. Gilley T.R. Tice (1990) ArticleTitleControlled vaccine release in the gut associated lymphoid tissues: orally administered biodegradable microspheres target the Peyer’s patches J Controlled Release 11 205–214 Occurrence Handle10.1016/0168-3659(90)90133-E
M. Fernandez-Alonso A. Rocha J.M. Coll (2001) ArticleTitleDNA vaccination by immersion and ultrasound to trout viral haemmorrhagic septicaemia virus Vaccine 19 3067–3075 Occurrence Handle10.1016/S0264-410X(01)00046-9
E. Hansen K. Fernandez G. Goldspink P. Butterworth P.K. Umeda K.-C. Chang (1991) ArticleTitleStrong expression of foreign genes following direct injection into fish muscle Fed Eur Biochem Soc 290 73–76
J. Heppell H.L. Davis (2000) Intramuscular injection of DNA vaccines in fish D.B. Lowrie R.G. Whalen (Eds) DNA Vaccines: Methods and Protocols Humana Press Totowa, N.J.
Herrera, A.A. 1996. Histology of Tilapia: Oreochromis niloticus. Quezon City, Philippines: Bureau of Fisheries and Aquatic Resources, Department of Agriculture.
K.W. Leong H.Q. Mao V.L. Truong-Le K. Roy S.M. Walsh J.T. August (1997) ArticleTitleDNA-polycation nanospheres as non-viral gene delivery vehicles J Controlled Release 53 183–193 Occurrence Handle10.1016/S0168-3659(97)00252-6
H.L. Robinson C.A.T. Torres (1997) ArticleTitleDNA vaccines Semin Immunol 9 271–283 Occurrence Handle10.1006/smim.1997.0083
K. Roy H-Q. Mao S-K. Huang K.W. Leong (1999) ArticleTitleOral gene delivery with chitosan-DNA nanoparticles generates immunologic protection in a murine model of peanut allergy Nat Med 5 387–391 Occurrence Handle10.1038/7385
J. Sambrook T. Maniatis E.F. Fritsch (1989) Molecular Cloning: A Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Press Cold Spring Harbor, N.Y.
H. Savas O. Güven (2001) ArticleTitleInvestigation of active substance release from poly (ethylene oxide) hydrogels Int J Pharm 224 151–158 Occurrence Handle10.1016/S0378-5173(01)00745-1
C.T. Sieu (2000) ArticleTitleSeeds of division Asiaweek 26 26–31
Sulaiman, Z.H., Chan, R.H.M., Simanjuntak, P.M., and Jami, Y.H.A. (2000). Gene transfer in food fish and prawn. Presented at the 7th ASEAN Food Conference, Manila, Philippines, November 19–22, 2000
R.A. Winegar J.A. Montefiore K.D. Suing K.G. O’Loughlin C.J. Rudd J.T. MacGregor (1996) ArticleTitleDetermination of tissue distribution of an intramuscular plasmid vaccine using PCR and in situ DNA hybridization Hum Gene Ther 7 2185–2194 Occurrence Handle10.1089/hum.1996.7.17-2185
R. Zeller (1997) Fixation, embedding, and sectioning of tissues, embryos, and single cells Ausubel Brent Kingston Moore Seidman Smith Struhl (Eds) Short Protocols in Molecular Biology John Wiley & Sons Mississauga, Ont
Acknowledgments
We thank Mr. Remus C. Villanueva for his invaluable support and indispensable role in making this project possible; Dr. Cynthia Nalo-Ochona, Ms. Hazel B. Alfon, Ms. Eloise Prieto, and Mr. Adrian Constantino for their technical assistance and support in the handling and staining of specimens; and Dr. Cynthia P. Saloma for the use of her microscope and camera.
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Ramos, E.A., Relucio, J.L.V. & Torres-Villanueva, C.A.T. Gene Expression in Tilapia Following Oral Delivery of Chitosan-Encapsulated Plasmid DNA Incorporated into Fish Feeds. Mar Biotechnol 7, 89–94 (2005). https://doi.org/10.1007/s10126-004-3018-0
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DOI: https://doi.org/10.1007/s10126-004-3018-0