The effectiveness of adenoviral vectors to deliver and express genes in rainbow trout, Oncorhynchus mykiss (Walbaum).
The efficacy of adenoviral vectors for gene delivery into fish cells, both in vitro and in vivo, was evaluated. Vectors utilized were of human adenovirus serotype 5 (Ad), which are commonly used in human clinical trials, but have not been assessed for gene delivery to fish. Because nothing is known about Ad receptors in fish, both an Ad (Ad5Luc1) with natural tropism for the coxsackie and adenovirus receptor (CAR), as well as an infectivity enhanced Ad (Ad5LucRGD) were included within this study. Gene expression was detected in cell lines using either vector. The levels seen with Ad5LucRGD were much higher than for Ad5Luc1 in most lines except CHSE-214. Transduction of CHSE-214 cells with Ad5Luc1 could be blocked with an excess of a competitive inhibitor, suggesting that these cells possess a CAR homologue thatmediates attachment of Ad, similar to that seen in mammalian cells. In vivo gene delivery was attempted by several methods, with significant expression seen only via intramuscular injection, although infection efficiency was low. Thus it was observed that several teleost cell lines are capable of being infected and one cell line expressed a human serotype adenoviral receptor homologue that aids in Ad infection. Additionally, in vivo studies indicated that muscle tissue of rainbow trout could be infected with Ad vectors, suggesting an alternative gene delivery strategy for this animal.
Overturf
K
K
USDA/ARS, Hagerman Fish Culture Experiment Station, Hagerman, ID 83332, USA. kennetho@uidaho.edu
LaPatra
S
S
Reynolds
P N
PN
D000260:Adenoviruses, Human / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D005393:Fish Diseases / Q000628:therapy*; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques / Q000662:veterinary*; D015316:Genetic Therapy / Q000662:veterinary*; D005822:Genetic Vectors / Q000592:standards; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000235:genetics*; D011991:Receptors, Virus / Q000235:genetics
D011991:Receptors, Virus; D008156:Luciferases