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Handmade cloned transgenic piglets expressing the nematode fat-1 gene

journal contribution
posted on 06.12.2017, 00:00 by P Zhang, Y Zhang, H Dou, J Yin, Y Chen, X Pang, Gabor VajtaGabor Vajta, L Bolund, Y Du, R Ma
Production of transgenic animals via somatic cell nuclear transfer (SCNT) has been adapted worldwide, but this application is somewhat limited by its relatively low efficiency. In this study, we used handmade cloning (HMC) established previously to produce transgenic pigs that express the functional nematode fat-1 gene. Codon-optimized mfat-1 was inserted into eukaryotic expression vectors, which were transferred into primary swine donor cells. Reverse transcriptase PCR (RT-PCR), gas chromatography, and chromosome analyses were performed to select donor clones capable of converting n-6 into n-3 fatty acids. Blastocysts derived from the clones that lowered the n-6/n-3 ratio to approximately 1:1 were transferred surgically into the uteri of recipients for transgenic piglets. By HMC, 37% (n = 558) of reconstructed embryos developed to the blastocyst stage after 7 days of culture in vitro, with an average cell number of 81 – 36 (n = 14). Three recipients became pregnant after 408 day-6 blastocysts were transferred into four naturally cycling females, and a total of 14 live offspring were produced. The nematode mfat-1 effectively lowered the n-6/n-3 ratio in muscle and major organs of the transgenic pig. Our results will help to establish a reliable procedure and an efficient option in the production of transgenic animals.

Funding

Other

History

Volume

14

Issue

3

Start Page

258

End Page

266

Number of Pages

9

eISSN

2152-4998

ISSN

2152-4971

Location

USA

Publisher

Mary Ann Liebert

Language

en-aus

Peer Reviewed

Yes

Open Access

No

External Author Affiliations

BGI Ark Biotechnology Co Ltd (Shenzen); Beijing ji yin zu yan jiu suo; TBA Research Institute; Zhongguo ke xue yuan; Zhongguo nong ye da xue;

Era Eligible

Yes

Journal

Cellular reprogramming.

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