[Construction of a tRNAVal promoter plasmid expressing shRNAs in mammalian cells which mediates RNA interference]
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Published on: August 18, 2010
Xiao-chun Lu1, Zhen-hua Yuan, Jian-qiang Peng
1Department of Geriatristic Cardiology, PLA General Hospital, Beijing 100853, China.
Objective:
Constructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference.
Methods:
A tRNAVal gene was amplified from human genomic DNA by PCR and replaced the last several bases of 3' end by a linker. The tRNAVal promoter after artificial mutation followed a shRNA sequence to luciferase was cloned into pUC18, Puc-tRNAVal, lucRi Cotransfected with pMAMneoLuc into BHK-21 cell to detect the effect of luciferase expression.
Results:
pUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%.
Conclusion:
The results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21.
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