Inhibition of Coxsackievirus B3 Replication by Small Interfering RNAs Requires Perfect Sequence Match in the Central Region of the Viral Positive Strand

P58IPKinhibits coxsackievirus-induced apoptosis via the PI3K/Akt pathway requiring activation of ATF6a and subsequent upregulation of mitofusin 2

“…Virus titres were determined by plaque assay as described previously (Yuan et al, 2005). Briefly, HeLa cells were seeded into 6-well plates (8 × 10 5 cells/well) and incubated at 37°C for 20 h to a confluence of approximately 90% and then washed with PBS and overlaid with 1 ml of virus-containing samples serially diluted in cell culture medium.…”

Progress in targeted delivery of siRNA to combat Coxsackievirus

“…Yuan and colleagues, for the first time, achieved 92% inhibition of Coxsackievirus B3 replication, synthesizing a siRNA targeting the CVB3 protease 2A gene. They demonstrated an inhibition of CVB3 replication in Hela cells using wild-type CVB3 virus together with employing an ongoing viral infection in a dose dependent manner for long term treatments (8-48 h) (Yuan et al, 2005). The author also reported the inhibition of CVB3 in HL-1 mouse cardiomyocyte cell lines for 48 h post infection at a MOI of 10, thereby showing that siRNA exhibits greatest potency in inhibiting CVB3 replication.…”

“…The 3′ UTR contains three stem-loops followed by a poly (A) tail. The genomic RNA can serve as a template for viral RNA transcription to synthesize more copies of positive genomic RNA through a negative-strand intermediate (Yuan et al, 2005). This viral genome encodes four capsid proteins VP1-VP4 and seven nonstructural proteins including two proteases 2A and 3C, and an RNA-dependent RNA polymerase 3D.…”

“…A relatively new approach for delivering siRNA is described in an earlier study by Yuan and colleagues and examines the effects of RNAi on CVB3 replication using five siRNAs targeting different regions of CVB3 genomic RNA (Yuan et al, 2005). Small interfering RNAs (siRNAs) are short, double stranded RNA (dsRNA) molecules that can target mRNA of a specific sequence for degradation via a cellular process known as RNA interference (RNAi) (Elbashir et al, 2001;Fire, et al, 1998;Vaucheret, et al, 2001;Baulcombe, 2002).…”

“…This result might be explained by the fact that the replicating negative strands of virus only exist as a double stranded form in the vesicles (Fields et al, 2001); thus, they are less likely to be accessible to siRNAs. Conversely, the positive strand is the recognition site for RNAi, which forms complementary base pairs with the antisense strand of the siRNA (Yuan et al, 2005). Based on this finding, the researcher suggests that the match between the positive strand of viral RNA and antisense strand of siRNA is critical for RNA interference.…”