[Construction and identification of matrix metalloproteinase inhibitor-1 siRNA eukaryotic expression vectors]

Zhonghua Gan Zang Bing Za Zhi. 2007 Mar;15(3):196-8.
[Article in Chinese]

Abstract

Objective: To construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.

Methods: The combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.

Results: Targeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.

Conclusion: We successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Gene Silencing
  • Genetic Vectors*
  • Hepatic Stellate Cells*
  • Plasmids
  • RNA, Small Interfering*
  • Rats
  • Tissue Inhibitor of Metalloproteinase-1 / genetics*
  • Transfection

Substances

  • RNA, Small Interfering
  • Tissue Inhibitor of Metalloproteinase-1