Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

  • Yuan, Yu-Guo (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Song, Shao-Zheng (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Zhu, Meng-Ming (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • He, Zheng-Yi (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Lu, Rui (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Zhang, Ting (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Mi, Fei (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Wang, Jin-Yu (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University) ;
  • Cheng, Yong (Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis/College of animal science and technology, Yangzhou University)
  • Received : 2016.09.19
  • Accepted : 2016.12.13
  • Published : 2017.08.01
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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 for BLG gene 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. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate biallelic 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.

Keywords

References

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