Silencing of African horse sickness virus VP7 protein expression in cultured cells by RNA interference

Stassen, Liesel; Huismans, H.; Theron, Jacques

doi:10.1007/s11262-007-0162-4

Cited by 2 publications

(2 citation statements)

References 41 publications

(38 reference statements)

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“…Moreover, clinical trials in humans were started using siRNA in the treatment of respiratory syncytial virus (RSV) infections (Haasnoot et al, 2007). For equine viruses, in vitro studies have shown that siRNA can reduce viral titers in cell cultures infected with equine arteritis virus (Heinrich et al, 2009), African horse sickness virus (Stassen et al, 2007) or Venezuelan equine encephalitis virus (O'Brien, 2007). More specifically for EHV-1, RNA interference has been effective both in vitro and in vivo, as assessed by a reduction in plaque formation in cell cultures and a reduction in clinical signs (weight loss) and viral replication within lung tissue in a murine model of EHV-1 infection (Fulton et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

The effect of siRNA treatment on experimental equine herpesvirus type 1 (EHV-1) infection in horses

et al. 2010

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Available vaccines fail to induce lasting and protective immunity to equine herpesvirus 1 (EHV-1) associated diseases. RNA interference is a novel approach showing promise for therapeutic use in outbreak situations. This study examined the effect of small interfering RNA (siRNA) on clinical signs as well as the presence of live virus and viral DNA in nasal secretions and peripheral blood mononuclear cells (PBMCs) in horses experimentally infected with EHV-1. siRNA targeting two EHV-1 genes (glycoprotein B and the origin binding protein) was administered 12h before and 12h after intranasal infection with EHV-1. Control horses received siRNA targeting firefly luciferase. A significantly smaller proportion (0/10) of horses receiving siRNA targeting viral genes required euthanasia due to intractable neurologic disease as compared to horses in the control group (3/4; p=0.01). There was no significant difference in the presence of live virus or viral DNA in the nasal secretions or PBMCs between the two groups. Future studies are necessary to define the relative contributions of host and virus factors in the development of the neurological form of the infection and to determine an optimal dosing regimen for metaphylactic or therapeutic use of siRNA for treating EHV-1 infection.

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Section: Introductionmentioning

confidence: 99%

The effect of siRNA treatment on experimental equine herpesvirus type 1 (EHV-1) infection in horses

et al. 2010

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“…On one hand, the method could be significantly improved by reducing this virus background with discriminating specific siRNAs. Very strong reduction of virus growth has been published for African horse sickness virus, another serogroup of the genus O rbivirus [20]. On the other hand, the amount of in vitro synthesized RNA in infected cells could be further increased to improve the efficiency to rescue reassortants.…”

Section: Resultsmentioning

confidence: 99%

Genetic modification of Bluetongue virus by uptake of "synthetic" genome segments

Gennip¹,

Veldman²,

Water³

et al. 2010

Virol J

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Since 1998, several serotypes of Bluetongue virus (BTV) have invaded several southern European countries. In 2006, the unknown BTV serotype 8 (BTV8/net06) unexpectedly invaded North-West Europe and has resulted in the largest BT-outbreak ever recorded. More recently, in 2008 BTV serotype 6 was reported in the Netherlands and Germany. This virus, BTV6/net08, is closely related to modified-live vaccine virus serotype 6, except for genome segment S10. This genome segment is closer related to that of vaccine virus serotype 2, and therefore BTV6/net08 is considered as a result of reassortment. Research on orbiviruses has been hampered by the lack of a genetic modification method. Recently, reverse genetics has been developed for BTV based on ten in vitro synthesized genomic RNAs. Here, we describe a targeted single-gene modification system for BTV based on the uptake of a single in vitro synthesized viral positive-stranded RNA. cDNAs corresponding to BTV8/net06 genome segments S7 and S10 were obtained by gene synthesis and cloned downstream of the T7 RNA-polymerase promoter and upstream of a unique site for a restriction enzyme at the 3'-terminus for run-off transcription. Monolayers of BSR cells were infected by BTV6/net08, and subsequently transfected with purified in vitro synthesized, capped positive-stranded S7 or S10 RNA from BTV8/net06 origin. "Synthetic" reassortants were rescued by endpoint dilutions, and identified by serotype-specific PCR-assays for segment 2, and serogroup-specific PCRs followed by restriction enzyme analysis or sequencing for S7 and S10 segments. The targeted single-gene modification system can also be used to study functions of viral proteins by uptake of mutated genome segments. This method is also useful to generate mutant orbiviruses for other serogroups of the genus Orbivirus for which reverse genetics has not been developed yet.

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Silencing of African horse sickness virus VP7 protein expression in cultured cells by RNA interference

Cited by 2 publications

References 41 publications

The effect of siRNA treatment on experimental equine herpesvirus type 1 (EHV-1) infection in horses

The effect of siRNA treatment on experimental equine herpesvirus type 1 (EHV-1) infection in horses

Genetic modification of Bluetongue virus by uptake of "synthetic" genome segments

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