Abstract
Using MLV (murine leukemia virus)-based retrovirus vectors encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein), we tried to make transgenic chickens carrying the transferred genes in their chromosomes. Twenty one days after virus injection beneath the blastoderms of unincubated chicken embryos (stage X, at laying), DNA isolated from the hatched chicks were analyzed by PCR with two sets of primers specific for EGFP (enhanced green fluorescence protein) gene or Neo R (E. coli neomycin resistant) gene. Among sixty-seven embryos injected with retrovirus, four of them were identified to carry the EGFP genes in their genomes. Remarkably, one transgenic chick showed presence of the retrovirus vector sequences in all organs differentiated from one of endoderm, mesoderm, and ectoderm. Expression of EGFP gene was not detected, however, the stable germ line transmission of transgene was verified in spermatozoa from the founder chicken and 50% of F 1 progenies.
Highlights
Compared to the transgenic mammals, the production of transgenic chicken has several advantages including shorter generation time, lower expense, fecundity, etc
One of the most critical factors in the successful production of transgenic chicken is an efficient gene transfer to the multiple blastodermal cells of the newly laid eggs designated as stage X (Eyal-Giladi and Kochav, 1976)
Until now most retrovirus vector systems used in the transgenic chicken production has been derived from avian retrovirus (Bosselman et al, 1990), while the virus vectors used in this study are based on MLV and designed to be packaged by VSV-G enabling the progeny viruses to infect most of vertebrate cells (Burns et al, 1993)
Summary
Compared to the transgenic mammals, the production of transgenic chicken has several advantages including shorter generation time, lower expense, fecundity, etc. Until now most retrovirus vector systems used in the transgenic chicken production has been derived from avian retrovirus (Bosselman et al, 1990), while the virus vectors used in this study are based on MLV (murine leukemia virus) and designed to be packaged by VSV-G (vesicular stomatitis virus glycoprotein) enabling the progeny viruses to infect most of vertebrate cells (Burns et al, 1993). Advantages of this pseudotyped hybrid retrovirus vector system include availability of highly concentrated virus stock and decreased possibility of homologous recombination between the vector and the dormant endogenous retrovirus in the target cell.
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