Document Detail


Handmade cloned transgenic piglets expressing the nematode fat-1 gene.
MedLine Citation:
PMID:  22686479     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Peng Zhang; Yidi Zhang; Hongwei Dou; Jingdong Yin; Yu Chen; Xinzhi Pang; Gabor Vajta; Lars Bolund; Yutao Du; Runlin Z Ma
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Publication Detail:
Type:  Clinical Trial; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cellular reprogramming     Volume:  14     ISSN:  2152-4998     ISO Abbreviation:  Cell Reprogram     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-06-12     Completed Date:  2012-10-25     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  101528176     Medline TA:  Cell Reprogram     Country:  United States    
Other Details:
Languages:  eng     Pagination:  258-66     Citation Subset:  IM    
Affiliation:
State Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
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MeSH Terms
Descriptor/Qualifier:
Adiposity / genetics,  physiology
Animals
Animals, Genetically Modified*
Body Constitution / genetics
Caenorhabditis elegans Proteins / genetics*,  metabolism,  physiology
Cells, Cultured
Cloning, Organism / methods,  veterinary*
Embryo Transfer
Fatty Acid Desaturases / genetics*,  metabolism,  physiology
Fatty Acids, Omega-3 / metabolism
Fatty Acids, Omega-6 / metabolism
Female
Lipid Metabolism / genetics,  physiology
Nematoda / genetics
Pregnancy
Swine* / genetics
Chemical
Reg. No./Substance:
0/Caenorhabditis elegans Proteins; 0/Fatty Acids, Omega-3; 0/Fatty Acids, Omega-6; 0/fat-1 protein, C elegans; EC 1.14.19.-/Fatty Acid Desaturases
Comments/Corrections

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