Efficient method for expressing transgenes in nonhuman primate embryos using a stable episomal vector

Transgenesis in the nonhuman primate can enhance the study of human biology by providing animal models for the study of primate‐specific physiology, pathophysiology, and embryonic development. Progress with this technology has been hindered by the inherent inefficiency of transgenesis, transgene sil...

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Bibliographic Details
Published in:Molecular reproduction and development 2002-05, Vol.62 (1), p.69-73
Main Authors: Wolfgang, Michael J., Marshall, Vivienne S., Eisele, Stephen G., Schotzko, Michele L., Thomson, James A., Golos, Thaddeus G.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Line
Epstein-Barr virus
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes, Reporter
Genetic Vectors
Green Fluorescent Proteins
Herpesvirus 4, Human - genetics
Humans
implantation
IVF
Luminescent Proteins - genetics
Macaca mulatta - embryology
Molecular and cellular biology
Molecular genetics
Plasmids
rhesus monkey
Transgenes
transgenic
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Summary:Transgenesis in the nonhuman primate can enhance the study of human biology by providing animal models for the study of primate‐specific physiology, pathophysiology, and embryonic development. Progress with this technology has been hindered by the inherent inefficiency of transgenesis, transgene silencing, and practical restrictions on the production of sufficient pronuclear stage nonhuman primate zygotes. We have developed a novel technique using an Epstein Barr virus (EBV)‐based episomal vector to produce rhesus monkey (Macaca mulatta) embryos expressing a transgene. Plasmid DNA containing the latent origin of replication, oriP, and Epstein Barr Nuclear Antigen‐1 (EBNA‐1) of EBV, as well as a CMV IE‐enhanced green fluorescent protein (eGFP) expression cassette, was introduced into rhesus embryos by direct pronuclear microinjection. We detected eGFP in early cleavage stage embryos (4–8 cell) and throughout the duration of culture (day 8–9 blastocysts) by epifluorescent microscopy. A 50% transduction rate was obtained with the EBV‐based vector. Microinjected embryos expressed eGFP and retained their developmental capacity as evidenced by development to the blastocyst stage. EBV‐based vectors present a novel and efficient means of delivering transgenes for the study of the molecular control of primate embryonic development. Mol. Reprod. Dev. 62: 69‐73, 2002. © 2002 Wiley‐Liss, Inc.
ISSN:1040-452X
1098-2795
DOI:10.1002/mrd.10059