Tetracycline 유도적인 RNAi System을 이용한 생쥐 성장 관련 유전자의 발현 억제
Inhibition of mIGF-1 and mGHR Gene Expression using Tetracycline-Inducible RNAi System in Mouse Liver Cell
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.
목차
ABSTRACT 서론 실험 방법 shRNA 합성 생쥐 shRNA를 도입한 Tet Vector System의 유전자 발현의 Knock-Down 효과 검정 결과 Tetracycline 유도적인 Retrovirus Vector System을 이용한 mIGF-1i의 Knock-Down 효과 검정 Tetracycline 유도적인 Retrovirus Vector System을 이용한 mGHRi의 Knock-Down 효과 검정 단백질 수준에서의 mIGF-1i와 mGHRi의 Knock-Down 효율 양상 확인 고찰 인용문헌