RNA interference inhibits hepatitis E virus mRNA accumulation and protein synthesis in vitro

Abstract

Hepatitis E virus (HEV) is a zoonotic pathogen to which several species, including human beings, pigs and rodents, are reported to be susceptible. To date, vaccines developed against HEV still need to be improved and a structural gene (ORF2), which encodes a capsid protein with high sequence conservation found across HEV genotypes, is a potential candidate. To exploit the possibility of using RNA interference (RNAi) as a strategy against HEV infection, four small interference RNA (siRNA) duplex targeting ORF2 gene were constructed. A challenge against HEV infection by RNAi was performed in A549 cells. Real-Time quantitative polymerase chain reaction (Real-Time qPCR) and Western blot assay demonstrated that four HEV specific siRNAs (si-ORF2-1, si-ORF2-2, si-ORF2-3 and si-ORF2-4) were capable of protecting cells against HEV infection with very high specificity and efficiency. The results suggest that RNAi is a potent anti-HEV infection prophylaxis strategy.

Molecular virology of hepatitis E virus

“…Mono- and di-hammerhead ribozymes designed against the 3′ end of the HEV genomic RNA were shown to inhibit expression from a reporter construct in HepG2 cells ( Sriram et al, 2003 ). In A549 cells infected with HEV, small interfering RNAs (siRNAs) against the orf2 region were shown to offer protection ( Huang et al, 2010 ). While such approaches are feasible in vitro , the delivery and targeting of such inhibitors in vivo would be the real challenge.…”

Plasmids Expressing shRNAs Specific to the Nucleocapsid Gene Inhibit the Replication of Porcine Deltacoronavirus In Vivo

“…For fighting viral diseases with RNAi, considerable attempts have been made to date. Previous reports demonstrated that the plasmid vector constructs containing shRNAs targeting CSFV [ 25 ], FMDV [ 26 ], TGEV [ 27 ], and PEDV [ 28 ] could effectively inhibit viral replications [ 35 , 36 , 37 , 38 ]. RNAi-based technology is becoming a potent therapeutic strategy to exert antiviral activity [ 39 ].…”

Molecular Virology of Hepatitis E Virus

“…Interference with HEV RNA replication has been attempted using ribozymes and small interfering RNAs. 123,124 Although such approaches are feasible in vitro, the delivery and targeting of such inhibitors in vivo would be the real challenge. Several recent reports show the efficacy of Ribavirin monotherapy in treating immunocompromised trans-plant patients with chronic HEV infection, [125][126][127] severe acute hepatitis E, 128 and acute on chronic liver failure.…”