Journal
TRANSGENIC RESEARCH
Volume 22, Issue 4, Pages 737-745Publisher
SPRINGER
DOI: 10.1007/s11248-012-9674-3
Keywords
Transgenic sheep; Lentiviral vectors; Replication competent lentivirus; Embryo microinjection
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Funding
- National Research Initiative Competitive Grant from the USDA National Institute of Food and Agriculture [2007-35205-17921]
- Biotechnology Risk Assessment Competitive Grant from the USDA National Institute of Food and Agriculture [2009-33120-20238]
- NHLBI National Gene Vector Biorepository [P40 HL024928]
- NIFA [581353, 2009-33120-20238] Funding Source: Federal RePORTER
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The safety of HIV-1 based vectors was evaluated during the production of transgenic sheep. Vectors were introduced into the perivitelline space of in vivo derived one-cell sheep embryos by microinjection then transferred into the oviducts of recipient females. At 60-70 days of gestation, a portion of the recipients were euthanized and tissues collected from both surrogates and fetuses. Other ewes were allowed to carry lambs to term. Inadvertent transfer of vector from offspring to surrogates was evaluated in 330 blood and tissue samples collected from 57 ewes that served as embryo recipients. Excluding uterine contents, none of the samples tested positive for vector, indicating that that the vector did not cross the fetal maternal interface and infect surrogate ewes. Evaluating ewes, fetuses and lambs for replication competent lentivirus (RCL); 84 serum samples analyzed for HIV-1 capsid by ELISA and over 600 blood and tissue samples analyzed by quantitative PCR for the VSV-G envelopes revealed no evidence of RCL. Results of these experiments provide further evidence as to the safety of HIV-1 based vectors in animal and human applications.
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