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http://dx.doi.org/10.12749/RDB.2014.38.3.99

Inhibition of mIGF-1 and mGHR Gene Expression using Tetracycline-Inducible RNAi System in Mouse Liver Cell  

Son, Hye Jin (Department of Physiology, Catholic University of Daegu School of Medicine)
Koo, Bon Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
Kwon, Mo Sun (Department of Physiology, Catholic University of Daegu School of Medicine)
Lee, Young Man (Department of Physiology, Catholic University of Daegu School of Medicine)
Kim, Teoan (Department of Physiology, Catholic University of Daegu School of Medicine)
Publication Information
Reproductive and Developmental Biology / v.38, no.3, 2014 , pp. 99-105 More about this Journal
Abstract
In this study, to further understand the mechanism of animal growth and to develop a miniature transgenic animal model, we constructed and tested tetracycline-inducible RNAi system using shRNA targeting the mRNA of mouse insulin-like growth factor (mIGF-1) or mouse growth hormone receptor (mGHR) gene. Quantitative real-time PCR analysis of mouse liver cell (Hepa1c1c7) cells transfected with these vectors showed 85% or 90% of expression inhibition effect of IGF-1 or GHR, respectively. In ELISA analysis, the protein level of IGF-1 in the cells expressing the shRNA targeting IGF-1 mRNA was reduced to 26% of non-transformed control cells. Unexpectedly, in case of using shRNA targeting GHR, the IGF-1 protein level was decreased to 75% of control cells. Further experiments are needed to explain the lower interference effect of GHR shRNA in IGF-1 protein. Accumulated knowledge of this approach could be applicable to a variety of related biological area including gene function study, gene therapy, development of miniature animals, etc.
Keywords
RNAi system; IGF-1; GHR; Albumin promoter; Tetracycline inducible gene expression;
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