A Small Interfering RNA Targeting Coxsackievirus B3 Protects Permissive HeLa Cells from Viral Challenge

Ahn, Jee-Yin; Jun, Eun Seok; Lee, Hui Sun; Yoon, Sun-Young; Kim, Dong Hou; Joo, Chul Hyun; Kim, Yoo Kyum; Lee, Heuiran

doi:10.1128/jvi.79.13.8620-8624.2005

Cited by 42 publications

(35 citation statements)

References 26 publications

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“…Among the numerous viruses that have been successfully silenced by RNAi, there are five representatives of the picornavirus family: poliovirus (19,20), human rhinovirus (45), enterovirus 71 (37), foot-and-mouth disease virus (7,9,36), and coxsackievirus B3 (1,40). Recently, we have shown that genome replication of an HAV replicon can be efficiently inhibited by small interfering RNAs (siRNAs) targeting either coding or noncoding regions of the genome (26,27,34).…”

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confidence: 99%

Silencing of Hepatitis A Virus Infection by Small Interfering RNAs

Kusov

Palmenberg

Sgro

et al. 2006

J Virol

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Infection by hepatitis A virus (HAV) can cause acute hepatitis and, rarely, fulminant liver failure, in particular in patients chronically infected with hepatitis C virus. Based on our previous observation that small interfering RNAs (siRNAs) can silence translation and replication of the firefly luciferase-encoding HAV replicon, we now exploited this technology to demonstrate the effect of siRNAs on viral infection in Huh-7 cells. Freshly and persistently infected cells were transfected with siRNAs targeting various sites in the HAV nonstructural genes. Compared to a single application, consecutive siRNA transfections targeting multiple sequences in the viral genome resulted in a more efficient and sustained silencing effect than a single transfection. In most instances, multiple applications of a single siRNA led to the emergence of viral escape mutants with mutated target sites that rendered these genomes resistant to RNA interference (RNAi). Efficient and sustained suppression of the viral infectivity was achieved after consecutive applications of an siRNA targeting a computer-predicted hairpin structure. This siRNA holds promise as a therapeutic tool for severe courses of HAV infection. In addition, the results provide new insight into the structural bases for sequencespecific RNAi.Hepatitis A virus (HAV) has a single-stranded RNA genome of 7.5 kb with positive polarity. As a member of the picornavirus family, it is unique in its biological properties, in particular, in its highly protracted replicative cycle in cell cultures and inability to shut off the host cell metabolism. HAV causes a self-limiting infection of the liver with usually mild or no symptoms in young patients, whereas adult patients might suffer from severe symptoms. In immune-compromised or chronically infected patients, fulminant hepatic failure is often associated with a high mortality rate (15, 52). Albeit vaccines against hepatitis A virus can prevent the disease (3, 14, 31), they are futile for fulminant hepatitis. Therefore, therapeutic intervention strategies to prevent progression to a life-threatening liver disease are still in need. Given that the rapid degradation of the HAV genome can ameliorate viral infection and thus prevent fulminant failure, RNA interference (RNAi) based on sequence-specific genome degradation may present a novel and specific therapeutic approach in preventing severe disease progression.Initially demonstrated in Caenorabditis elegans and Drosophila melanogaster, RNAi is now recognized as a valuable tool to silence viral infections (reviewed in references 22, 48, and 50). Among the numerous viruses that have been successfully silenced by RNAi, there are five representatives of the picornavirus family: poliovirus (19,20), human rhinovirus (45), enterovirus 71 (37), foot-and-mouth disease virus (7, 9, 36), and coxsackievirus B3 (1, 40). Recently, we have shown that genome replication of an HAV replicon can be efficiently inhibited by small interfering RNAs (siRNAs) targeting either coding or noncoding regions...

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confidence: 99%

Silencing of Hepatitis A Virus Infection by Small Interfering RNAs

Kusov

Palmenberg

Sgro

et al. 2006

J Virol

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“…A number of compounds designed to inhibit picornavirus infections through an antisense-mediated mechanism have generated positive preclinical results but are still early in the development process. Antisense phophorothioate DNA has been used against coxsackievirus type B3 (CVB3) in vitro (57) and in vivo (62), antisense RNA has been used against foot-and-mouth disease virus (45), and small interfering RNA has been used against several picornaviruses (3,20,21,26,42,47), all with some success. Phosphorodiamidate morpholino oligomers (PMO) are composed of individual subunits consisting of a DNA base connected to a morpholine ring and phophorodiamidate-linked backbone (50).…”

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confidence: 99%

A Morpholino Oligomer Targeting Highly Conserved Internal Ribosome Entry Site Sequence Is Able To Inhibit Multiple Species of Picornavirus

Stone

Rijnbrand

Stein

et al. 2008

Antimicrob Agents Chemother

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“…Other groups, however, did not observe such effects. For example, Ahn et al (2005) did not find additive antiviral effects of 2 shRNAs against CVB-3, and Koller et al (2006) even reported that 2 siRNAs may compete for RNA-induced silencing complex (RISC) binding and thereby reduce each other's activity. We carried out additional experiments and found the inhibitory activity of an antiviral siRNA to decrease upon addition of a second (unspecific) siRNA (data not shown).…”

Section: Discussionmentioning

confidence: 99%

“…In mammalian cells RNAi can be induced by transient treatment with small interfering RNAs (siRNAs) or by long-term and stable expression using promoter-driven short hairpin RNAs (shRNAs). Several groups have employed RNAi to inhibit viruses, for example, the human immunodeficiency virus (Berkhout, 2009), influenza virus (Suzuki et al, 2009), and coxsackievirus B3 (Ahn et al, 2005;Merl et al, 2005). Viruses often mutate their target sequence, however, to escape RNAi treatment.…”

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confidence: 99%