Evaluating the potential of germ line transmission after intravenous administration of recombinant adenovirus in the C3H mouse.
The goal of this study is to assess the likelihood that an adenoviral vector disseminated to gonads will be transmitted to offspring. This study is based on the observation that systemically administered vector can be detected in both ovaries and testes, using sensitive nested PCR techniques. Although the extent of vector dissemination to gonads is extremely small, as it is detectable only by nested PCR, it is unclear where it is located within these tissues and whether the DNA is capable of integration and transmission to offspring. A protocol was developed in C3H mice to address this question. Both male and female C3H mice were injected with a high dose of H5.001CBhOTC, an E1- and E4-deleted vector expressing human ornithine transcarbamylase. This dose of vector was sufficient to target 80% of hepatocytes (Gao et al., J. Virol. 1996; 70:8934-8943) and disseminate, at low levels, to both ovaries and testes in 94% of animals as determined by PCR. Vector-administered animals and controls were mated and 814 offspring were evaluated for germ line transmission of the adenoviral vector by DNA hybridization of total cellular DNA extracted from the fetus. Southern blot analysis showed no evidence of germ line transmission in 578 offspring of crosses in which either one or both parents received recombinant adenovirus.
Institute for Human Gene Therapy, Department of Molecular and Cellular Engineering, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
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Ye
G P
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Gao
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Pabin
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Raper
J M
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Wilson
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005333:Fetus / Q000201:enzymology / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D005854:Germ Cells / Q000201:enzymology*; D006066:Gonads / Q000821:virology*; D006801:Humans; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009954:Ornithine Carbamoyltransferase / Q000235:genetics; D016133:Polymerase Chain Reaction
D004279:DNA, Viral; D009954:Ornithine Carbamoyltransferase