title: TALEN-mediated modification of the bovine genome for large-scale production of human serum albumin.
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.
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Winston-Salem, North Carolina, United States of America.
authors.forename: Shaida
authors.initials: S
authors.lastname: Moghaddassi
authors.affiliation: Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, United States of America.
authors.forename: Will
authors.initials: W
authors.lastname: Eyestone
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Winston-Salem, North Carolina, United States of America.
authors.forename: Colin E
authors.initials: CE
authors.lastname: Bishop
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000235:genetics; D002478:Cells, Cultured; D003851:Deoxyribonucleases / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005818:Genetic Engineering; D016678:Genome; D006801:Humans; D008099:Liver / Q000378:metabolism; D008297:Male; D008892:Milk / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D012709:Serum Albumin / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D012709:Serum Albumin; D003851:Deoxyribonucleases