Construction and identification of matrix metalloproteinase inhibitor-1 siRNA eukaryotic expression vectors / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 196-198, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-285431
ABSTRACT
<p><b>OBJECTIVE</b>To construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>Targeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.</p><p><b>CONCLUSION</b>We successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Transfection
/
Cell Line
/
Tissue Inhibitor of Metalloproteinase-1
/
Gene Silencing
/
RNA, Small Interfering
/
Hepatic Stellate Cells
/
Genetic Vectors
/
Genetics
Type of study:
Diagnostic study
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2007
Type:
Article
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