In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics.

In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics. Research Abstract Details 

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In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics. Abstract Text:

Allison Groseth,Thomas Hoenen,Judie B Alimonti,Florian Zielecki,Hideki Ebihara,Steven Theriault,Ute Stroher,Stephan Becker,Heinz Feldmann,Allison Groseth,Thomas Hoenen,Judie B Alimonti,Florian Zielecki,Hideki Ebihara,Steven Theriault,Ute ,Stephan Becker,Heinz Feldmann,Allison Groseth,Thomas Hoenen,Judie B Alimonti,Florian Zielecki,Hideki Ebihara,Steven Theriault,Ute ,Stephan Becker,Heinz Feldmann,

Background. Recent reports indicate the possibility of using small interfering RNAs (siRNAs) to treat filovirus infections; however, they also show that the effectiveness of this approach is highly dependent on target site selection. Therefore, we explored the application of minigenomes as screening tools to identify functional siRNA targets under biosafety level 2 conditions.Methods. siRNA candidates were screened using the minigenome system to identify those with potential antiviral activity, compared with controls with poor predicted function on the basis of design guidelines, or those that were noncomplementary to Zaire ebolavirus (ZEBOV). These findings were then validated in cell culture by use of a previously developed ZEBOV expressing green fluorescent protein (ZEBOV-GFP), which allowed siRNA function to be easily assessed via flow cytometry or focus formation.Results. The most promising siRNA based on minigenome screening, targeting the nucleoprotein (NP) mRNA (ZNP1), also reduced protein expression and decreased viral titers after infection with ZEBOV-GFP to an extent similar to that reported for an siRNA recently shown to be therapeutic in guinea pigs.Conclusions. Minigenome screening appears to be an effective and convenient method of evaluating the therapeutic potential of siRNA targets, and findings suggest that its use would increase success rates in later stages of siRNA testing.

In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics. Publishing Authors By Initials

A Groseth,T Hoenen,JB Alimonti,F Zielecki,H Ebihara,S Theriault,U Stroher,S Becker,H Feldmann,A Groseth,T Hoenen,JB Alimonti,F Zielecki,H Ebihara,S Theriault,U ,S Becker,H Feldmann,A Groseth,T Hoenen,JB Alimonti,F Zielecki,H Ebihara,S Theriault,U ,S Becker,H Feldmann,

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In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: The Journal of infectious diseases

VOLUME: 196 Suppl 2

Page Numbers: S382-9

Journal Abbreviation: J. Infect. Dis.

ISSN: 0022-1899

DAY: 15

MONTH: Nov

YEAR: 2007

In Vitro Evaluation of Antisense RNA Efficacy against Filovirus Infection, by Use of Reverse Genetics. Information

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LANGUAGE: eng

NlmUniqueID: 413675

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AFFILIATION: National Laboratory for Zoonotic Diseases and Special Pathogens, National Microbiology Laboratory, Public Health Agency of Canada, Manitoba, R3E 3R2, Canada. Heinz_Feldmann@phac-aspc.gc.ca.

Country: United States

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MEDLINETA: J Infect Dis

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