Transgenic sheep generated by lentiviral vectors: safety and integration analysis of surrogates and their offspring

Transgenic Research - Tập 22 - Trang 737-745 - 2012
Kenneth Cornetta1, Kimberly Tessanne2, Charles Long2, Jing Yao1, Carey Satterfield3, Mark Westhusin2
1Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, USA
2Departments of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, USA
3Department of Animal Sciences, Texas A&M University, College Station, USA

Tóm tắt

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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