Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer
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The production of animals by somatic cell nuclear transfer (SCNT) is inefficient, with approximately 2 % of micromanipulated oocytes going to term and resulting in live births. However, it is the most commonly used method for the generation of cloned transgenic livestock as it facilitates the attainment of transgenic animals once the nuclear donor cells are stably transfected and more importantly as alternatives methods of transgenesis in farm animals have proven even less efficient. Here we describe piggyBac-mediated transposition of a transgene into porcine primary cells and use of these genetically modified cells as nuclear donors for the generation of transgenic pigs by SCNT. Gene transfer by piggyBac transposition serves to provide an alternative approach for the transfection of nuclear donor cells used in SCNT.
KeywordspiggyBac Transposon Transposase SCNT Cloning Transgenesis
This study was supported by a grant from the National Science Foundation for Young Scholars of China (Grant Number: 31101689), a grant from the National High Technology Research and Development Program of China (863 Program, Grant Number: 2011AA100304), a grant from Department of Science and Technology of Guangdong, China (Grant Number: 2011A020901001) and by National Institutes of Health Grants 5P20RR024206 and R01 GM083158-01A1 (to S.M.).
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