Specific inhibition of Raf-1 gene expression in HNE1 by RNA interference / 中华耳鼻咽喉头颈外科杂志
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
;
(12): 805-808, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-315591
ABSTRACT
<p><b>OBJECTIVE</b>To silence the expression of Raf-1 gene in HNE1 cells using vector-based RNA interference (RNAi) technique.</p><p><b>METHODS</b>The vector containing the human U6 promoter was used for targeted gene silencing when a dsDNA oligonucleotide encoding an appropriate shRNA was ligated into the vector, and 67nt oligonucleotide fragment was inserted into the downstream of the U6 promoter. Plasmids containing different Raf-1 target sequences [ (1) pshuttle-Raf-1-a( 225), (2) pshattle-Raf-1-b ( 358) and (3) pshuttle-Raf-1-c(474)], were transfected into HNE1 cells. Expression of Raf-1 mRNA was assayed by RT-PCR. Apoptosis were determined by cytometry.</p><p><b>RESULTS</b>Vector-based RNAi had advantages over antisense RNA because it could be delivered to the target cell more efficiently, and effect could last longer. Raf-1 expression could be inhibited by plasmid-expressed shRNA. Three different targeting sequences were selected from Raf-1 gene, and the inhibitory effect of pSIREN shuttle-Raf-1-b (358) was biggest.</p><p><b>CONCLUSION</b>Raf-1 expression in HNE1 cells can be inhibited significantly using plasmid-based RNAi.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
RNA, Messenger
/
Transfection
/
Gene Expression
/
Nasopharyngeal Neoplasms
/
Promoter Regions, Genetic
/
Apoptosis
/
Proto-Oncogene Proteins c-raf
/
RNA Interference
/
Cell Line, Tumor
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
Year:
2006
Type:
Article
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