Induction of antiviral RNA interference by influenza A virus in human cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE89514 | GEO | 2016/11/07
SECONDARY ACCESSION(S): PRJNA352417
REPOSITORIES: GEO
Similar Datasets
Project description:Plants and invertebrates protect themselves from viruses through RNA interference (RNAi), yet it remains unknown whether this defense mechanism exists in mammals. Antiviral RNAi involves the processing of viral long double-stranded (ds) RNA molecules into small interfering RNAs (siRNAs) by the ribonuclease (RNAse) III Dicer. These siRNAs are incorporated into effector complex(es) containing members of the Argonaute (Ago) protein family and guide silencing of complementary target viral RNAs. Here, we detect the accumulation of phased Dicer-dependent virus-derived siRNA (viRNAs) and demonstrate their loading into Ago2 after infection of mouse embryonic stem (ES) cells with Encephalomyocarditis virus (EMCV). We further show that the production of these viRNAs is drastically reduced, yet not completely abolished, if ES cells are first induced to differentiate before infection. Finally, we reveal that the mammalian virus Nodamura virus (NoV) encodes for a protein that counteracts such antiviral RNAi in ES cells supporting the existence of an effective RNAi-based immunity in mammals. Infection of wild-type or mutant mouse ES cells and analysis of small RNAs from total extracts or immunoprecipitated components of the RNAi pathway
2013-10-10 | E-GEOD-43153 | biostudies-arrayexpress
Project description:Plants and invertebrates protect themselves from viruses through RNA interference (RNAi), yet it remains unknown whether this defense mechanism exists in mammals. Antiviral RNAi involves the processing of viral long double-stranded (ds) RNA molecules into small interfering RNAs (siRNAs) by the ribonuclease (RNAse) III Dicer. These siRNAs are incorporated into effector complex(es) containing members of the Argonaute (Ago) protein family and guide silencing of complementary target viral RNAs. Here, we detect the accumulation of phased Dicer-dependent virus-derived siRNA (viRNAs) and demonstrate their loading into Ago2 after infection of mouse embryonic stem (ES) cells with Encephalomyocarditis virus (EMCV). We further show that the production of these viRNAs is drastically reduced, yet not completely abolished, if ES cells are first induced to differentiate before infection. Finally, we reveal that the mammalian virus Nodamura virus (NoV) encodes for a protein that counteracts such antiviral RNAi in ES cells supporting the existence of an effective RNAi-based immunity in mammals.
2013-10-10 | GSE43153 | GEO
Project description:While the issue of whether RNA interference (RNAi) ever forms part of the antiviral innate immune response in mammalian somatic cells remains controversial, there is considerable evidence demonstrating that few, if any, viral small interfering RNAs (siRNAs) are produced in infected cells. Moreover, total inhibition of RNAi by mutational inactivation of key RNAi factors, such as Dicer or Argonaut 2, has been shown to not enhance virus replication. One potential explanation for this lack of inhibitory effect is that mammalian viruses encode viral suppressors of RNAi (VSRs) that are so effective that viral siRNAs are not produced in infected cells. Indeed, a number of mammalian VSRs have been described of which the most prominent is the influenza A virus (IAV) NS1 protein, which has not only been reported to inhibit RNAi in plants and insects but also to prevent the production of viral siRNAs in IAV-infected human cells. Here, we confirm that an IAV mutant that lacks the viral NS1 protein indeed differs from wild type IAV in that it induces the production of readily detectable levels of Dicer-dependent viral siRNAs in infected human cells. However we also demonstrate that these siRNAs have little if any inhibitory effect on IAV gene expression.
2018-07-03 | GSE111572 | GEO
Project description:In vertebrates, the presence of viral RNA in the cytosol is sensed by members of the RIG-I like receptor (RLR) family , which signal to induce production of type I interferons (IFN). These key anti-viral cytokines act in a paracrine and autocrine manner to induce hundreds of interferon-stimulated genes (ISGs), whose protein products restrict viral entry, replication and budding. ISGs include the RLRs themselves: RIG-I, MDA5 and the least-studied family member, LGP2. In contrast, the IFN system is absent in plants and invertebrates, which defend themselves from viral intruders using RNA interference (RNAi). In RNAi, the endoribonuclease Dicer cleaves virus-derived double stranded RNA (dsRNA) into small interfering RNAs (siRNAs) that target complementary viral RNA for cleavage. Interestingly, the RNAi machinery is conserved in mammals and we have recently demonstrated that it is able to participate in mammalian antiviral defence in conditions in which the IFN system is suppressed. In contrast, when the IFN system is active, one or more ISGs act to mask or suppress antiviral RNAi. Here, we demonstrate that LGP2 constitutes one of the ISGs that can inhibit antiviral RNAi in mammals. We identify Dicer as an LGP2-associated protein and show that LGP2 inhibits Dicer cleavage of dsRNA into siRNAs both in vitro and in vivo. Further, we show that in cells lacking an IFN response, ectopic expression of LGP2 interferes with RNAi-dependent suppression of gene expression. Thus, the inefficiency of RNAi as a mechanism of antiviral defence in mammalian somatic cells can be in part attributed to Dicer inhibition by LGP2 induced by type I IFNs.
2018-01-17 | PXD008364 | Pride
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE242867 | GEO
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE242869 | GEO
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE242868 | GEO
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE242866 | GEO
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE242870 | GEO
Project description:Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi has different functions in eukaryotes including gene regulation, antiviral innate immunity or defense against transposable elements. In mammals, RNAi is constrained by inefficient cleavage of dsRNA by Dicer because it is adapted to produce microRNAs, a different class of small RNAs. An exception of the rule is highly active RNAi in mouse oocytes, which employs a truncated Dicer isoform (ΔHEL1). A homozygous mutation of murine Dicer to express only the truncated variant causes major dysregulation of microRNAs and perinatal lethality. Here, we report the phenotype and RNAi activity in DicerΔHEL1/wt mice, which are viable, fertile, slightly smaller, and show minimal miRNome changes. At the same time, endogenous siRNA levels are increased by an order of magnitude in DicerΔHEL1/wt mice. We show that siRNA abundance is limited by available dsRNA but not by the Protein Kinase R, an innate immunity factor shown to limit siRNA biogenesis in cultured cells. Using dsRNA expressed from a transgene, functional RNAi in vivo was successfully induced in the heart. DicerΔHEL1/wt mice thus represent a new model for researching mammalian canonical RNAi in vivo and offer an unprecedented platform for addressing earlier claims about its biological roles.
2024-04-08 | GSE259395 | GEO