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10.3390/molecules15107197

http://scihub22266oqcxt.onion/10.3390/molecules15107197
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20956884!6259191!20956884
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suck abstract from ncbi


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pmid20956884      Molecules 2010 ; 15 (10): 7197-207
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  • 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 PMID20956884show 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]
  • |Sequence Alignment[MESH]
  • |Severe acute respiratory syndrome-related coronavirus/*genetics[MESH]


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