Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Inhibition of mIGF-1 and mGHR Gene Expression using Tetracycline-Inducible RNAi System in Mouse Liver Cell

Tetracycline 유도적인 RNAi System을 이용한 생쥐 성장 관련 유전자의 발현 억제

  • 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)
  • 손혜진 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 구본철 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 권모선 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 이영만 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 김태완 (대구가톨릭대학교 의과대학 생리학교실)
  • Received : 2014.08.25
  • Accepted : 2014.09.16
  • Published : 2014.09.30
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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

References

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