Generation of transgenic goats by pronuclear microinjection: a retrospective analysis of a commercial operation (1995–2012)
Production of transgenic founder goats involves introducing and stably integrating an engineered piece of DNA into the genome of the animal. At LFB USA, the ultimate use of these transgenic goats is for the production of recombinant human protein therapeutics in the milk of these dairy animals. The transgene or construct typically links a milk protein specific promoter sequence, the coding sequence for the gene of interest, and the necessary downstream regulatory sequences thereby directing expression of the recombinant protein in the milk during the lactation period. Over the time period indicated (1995–2012), pronuclear microinjection was used in a number of programs to insert transgenes into 18,120, 1- or 2- cell stage fertilized embryos. These embryos were transferred into 4180 synchronized recipient females with 1934 (47%) recipients becoming pregnant, 2594 offspring generated, and a 109 (4.2%) of those offspring determined to be transgenic. Even with new and improving genome editing tools now available, pronuclear microinjection is still the predominant and proven technology used in this commercial setting supporting regulatory filings and market authorizations when producing founder transgenic animals with large transgenes (> 10 kb) such as those necessary for directing monoclonal antibody production in milk.
KeywordsTransgenic Goat Microinjection Recombinant therapeutics
The authors would like to thank Charles R. Long, Ph.D., Associate Professor, Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas for his critical review of this manuscript. The authors would also like to thank Xiuchun (Cindy) Tian, Ph.D., Professor of Biotechnology, Department of Animal Science, University of Connecticut, Storrs, Connecticut for her critical review of the data and recommendations on the appropriate application of statistical analyses.
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