title: Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases.
abstract: OBJECTIVE
  To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases.
  
  
  METHODS
  TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT).
  
  
  RESULTS
  The targeted efficiency forBLGgene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d.
  
  
  CONCLUSIONS
  This finding signifies the combined use of TALENs and SCNT can generate bi-allelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.|Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou, 225001, China.
authors.forename: Yu-Guo
authors.initials: YG
authors.lastname: Yuan
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Shao-Zheng
authors.initials: SZ
authors.lastname: Song
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Meng-Ming
authors.initials: MM
authors.lastname: Zhu
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Zheng-Yi
authors.initials: ZY
authors.lastname: He
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Lu
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Mi
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Jin-Yu
authors.initials: JY
authors.lastname: Wang
authors.affiliation: Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.|Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou, 225001, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Cheng
keywords: Beta-lactoglobulin; Gene Targeted; Goat; Human Lacotoferrin; Nuclear Transfer; Transcription Activator-Like Effector Nucleases