1. TALEN-mediated modification of the bovine genome for large-scale production of human serum albumin.
- Author
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Moghaddassi S, Eyestone W, and Bishop CE
- Subjects
- Animals, Animals, Genetically Modified, Blastocyst physiology, Cattle genetics, Cells, Cultured, Deoxyribonucleases genetics, Female, Fibroblasts metabolism, Genome, Humans, Liver metabolism, Male, Milk metabolism, Nuclear Transfer Techniques, Serum Albumin genetics, Transfection, Genetic Engineering, Serum Albumin biosynthesis
- Abstract
As an initial step towards creating genetically modified cattle as a biopharming source of recombinant human serum albumin (rHSA), we report modification of the bovine albumin (bA) locus by transcription activator-like effector nuclease (TALEN)-stimulated homology-directed repair (HDR). Pedigreed bovine fibroblasts were co-transfected with TALENs and an 11.5-kb human serum albumin (HSA) minigene donor construct, designed to simultaneously disrupt and replace bovine serum albumin (BSA) expression with controlled rHSA expression in both the liver and the milk. Targeted integration of the HSA minigene was confirmed in transfected fibroblasts at a frequency of approximately 11% and transgenic bovine embryos were produced from targeted fibroblasts using somatic cell nuclear transfer (SCNT). The research delineated here lays the foundation for the future generation of transgenic rHSA cattle with the potential to provide a large-scale, reliable, and quality-controlled source of rHSA.
- Published
- 2014
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