Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk

• DOI: 10.1038/srep10482
• Publication Date: 2015-05-21T11:23:09Z
• Authors: Chenchen Cui; Yujie Song; Jun Liu; Hengtao Ge; Qian Li; Hui Huang; Linyong Hu; Hongmei Zhu; Yaping Jin; Yong Zhang
• Keywords: Gene targeting; Gene knockout; Lactoferrin; Zinc finger nuclease

Abstract β-Lactoglobulin (BLG) is a major goat’s milk allergen that absent in human milk. Engineered endonucleases, including transcription activator-like effector nucleases (TALENs) and zinc-finger nucleases, enable targeted genetic modification livestock. In this study, TALEN-mediated gene knockout followed by knock-in were used to generate BLG goats as mammary gland bioreactors for large-scale production of lactoferrin (hLF). We introduced precise modifications the goat genome at frequencies approximately 13.6% 6.09% first second sequential targeting, respectively, using targeting vectors underwent TALEN-induced homologous recombination (HR). Analysis from cloned revealed hLF expression or/and decreased levels heterozygous well absence homozygous goats. Furthermore, events somatic cells can be transmitted through germline after SCNT. Our result suggests via HR may expedite genetically engineered livestock agriculture biomedicine.