Preimplantation screening for transgenesis using an embryonic specific promoter and green fluorescent protein

Cloning
J T KeiserR L Page

Abstract

We report enrichment in the efficiency of generating mice transgenic for expression of a human protein in their milk using GFP-mediated preimplantation screening. The transgene array consisted of a functional gene (human alpha-1 antitrypsin under the control of the ovine BLG promoter) linked 5' to a reporter gene (GFP under the control of the murine Oct-4 promoter). GFP expression was detected in blastocysts by fluorescence microscopy and green and nongreen embryos were transferred to recipients in separate groups. In the first experiment, of seven pups that resulted from the transfer of blastocysts expressing GFP, five (71%) were transgenic. The experiment was repeated and of 12 pups that resulted from transfer of GFP-expressing blastocysts, 11 were transgenic (92%). The presence of the reporter cassette used for preimplantation screening did not affect the expression level of alpha-1-antitrypsin in the milk of the transgenic mice. In addition, in a related experiment wherein the GFP reporter gene was co-injected with a second mammary-specific transgene, pINC, no effect on transgene expression was observed. For mice transgenic for the mammary-specific gene alone, expression levels for four different lines were 192, 197, 382, a...Continue Reading

References

Dec 1, 1992·Molecular Reproduction and Development·T G Burdon, R J Wall
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Dec 1, 1994·Molecular Reproduction and Development·C CousensG T O'Neill
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Citations

Jun 5, 2008·Transgenic Research·Elena PopovaAlexander Krivokharchenko
Jan 7, 2004·Biochemical and Biophysical Research Communications·Vikram DevganPolani B Seshagiri
Oct 23, 2002·Biology of Reproduction·Shu-Hung ChenDavid L Ayares
Jan 22, 2003·Luminescence : the Journal of Biological and Chemical Luminescence·Yong A YuKerby Oberg
Dec 31, 2010·Journal of Virology·M R HajimoradM A Saghai Maroof
Dec 17, 2010·Journal of Virology·Jochen BodemAxel Rethwilm
Aug 29, 2006·PLoS Pathogens·Sacha LucchiniJay C D Hinton

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