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2010 ; 15
(10
): 7197-207
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
Small interfering RNA effectively inhibits the expression of SARS coronavirus
membrane gene at two novel targeting sites
#MMPMID20956884
Wang Y
; Cao YL
; Yang F
; Zhang Y
; Wang SH
; Liu L
Molecules
2010[Oct]; 15
(10
): 7197-207
PMID20956884
show ga
Small interfering RNA (siRNA) is a class of duplex RNA molecules of 21-25 nt
nucleotides in length functioning post-transcriptionally to downregulate targeted
gene expression. The membrane (M) protein of severe acute respiratory
syndrome-associated coronavirus (SARS-CoV) is highly abundant during viral
infections and is a critical element for viral assembly. Nucleotide substitution
in the viral genome occurs frequently during SARS-CoV infection. In the current
study, we analyzed the M gene sequences derived from 15 SARS-CoV isolates and
uncovered six nucleotide substitutions among these isolates. Interestingly, these
nucleotide substitutions are all located at the 5' half of the M gene. Based on
this information and previous reports, we created two novel siRNAs targeting two
unexplored and well conserved regions in the M gene. The effects of these two
siRNAs were tested by semi-quantitative RT-PCR and EGFP-M fusion gene expression.
The results demonstrated that both siRNAs effectively and specifically blocked
the targeted gene expression. Real time quantitative RT-PCR (qRT-PCR) revealed
that siRNA targeting the 3' half of the M gene (si-M2) induced more potent
inhibition than that targeting the 5' half (si-M1). Both si-M1 and si-M2
significantly downregulated M gene mediated upregulation of interferon b
expression. Thus, our results indicate that SARS-CoV M gene specific siRNA might
function in a sequence-dependent manner.
|*Genes, Viral
[MESH]
|*RNA Interference
[MESH]
|Base Sequence
[MESH]
|Coronavirus M Proteins
[MESH]
|HEK293 Cells
[MESH]
|Humans
[MESH]
|Molecular Sequence Data
[MESH]
|RNA, Small Interfering/genetics/*metabolism
[MESH]