Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 737-745 |
| Number of pages | 9 |
| Journal | Transgenic Research |
| Volume | 22 |
| Issue number | 4 |
| DOIs | |
| State | Published - Aug 2013 |
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Keywords
- Embryo microinjection
- Lentiviral vectors
- Replication competent lentivirus
- Transgenic sheep
ASJC Scopus subject areas
- Biotechnology
- Genetics
- Agronomy and Crop Science
- Animal Science and Zoology
Cite this
Transgenic sheep generated by lentiviral vectors : Safety and integration analysis of surrogates and their offspring. / Cornetta, Kenneth; Tessanne, Kimberly; Long, Charles; Yao, Jing; Satterfield, Carey; Westhusin, Mark.
In: Transgenic Research, Vol. 22, No. 4, 08.2013, p. 737-745.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transgenic sheep generated by lentiviral vectors
T2 - Safety and integration analysis of surrogates and their offspring
AU - Cornetta, Kenneth
AU - Tessanne, Kimberly
AU - Long, Charles
AU - Yao, Jing
AU - Satterfield, Carey
AU - Westhusin, Mark
PY - 2013/8
Y1 - 2013/8
N2 - 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.
AB - 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.
KW - Embryo microinjection
KW - Lentiviral vectors
KW - Replication competent lentivirus
KW - Transgenic sheep
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UR - http://www.scopus.com/inward/citedby.url?scp=84880513809&partnerID=8YFLogxK
U2 - 10.1007/s11248-012-9674-3
DO - 10.1007/s11248-012-9674-3
M3 - Article
VL - 22
SP - 737
EP - 745
JO - Transgenic Research
JF - Transgenic Research
SN - 0962-8819
IS - 4
ER -