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siRNAs pools generated in Escherichia coli exhibit strong RNA-interference activity against influenza virus genomic sequences
Virology. 2022 Dec 31;579:38-45. doi: 10.1016/j.virol.2022.12.013. Online ahead of print.ABSTRACTDue to the recurrent pandemic outbreaks that occurred during the last century, Influenza A viruses are considered a serious potential danger to human health. Among the innate immune pathways in eukaryotes, RNA interference plays a significant role in the interaction between viruses and host cells. RNA interference is addressed by small dsRNA molecules produced by the host itself (miRNAs, i.e. "micro-RNAs") but can be triggered also by the administration of exogenous short RNAs (siRNAs, "short interfering RNAs"). In this work, a...
Source: Virology - January 4, 2023 Category: Virology Authors: Riccardo Villa Sabrina Renzi Silvia Dotti Franco Lucchini Source Type: research

CDC25B promotes influenza A virus replication by regulating the phosphorylation of nucleoprotein.
Abstract Cell division cycle 25 B (CDC25B) is a member of the CDC25 phosphatase family. It can dephosphorylate cyclin-dependent kinases and regulate the cell division cycle. Moreover, siRNA knockdown of CDC25B impairs influenza A virus (IAV) replication. Here, to further understand the regulatory mechanism of CDC25B for IAV replication, a CDC25B-knockout (KO) 293T cell line was constructed using CRISPR/Cas9. The present data indicated that the replication of IAV was decreased in CDC25B-KO cells. Additionally, CDC25B deficiency damaged viral polymerase activity, nucleoprotein (NP) self-oligomerization, and NP nucle...
Source: Virology - September 18, 2018 Category: Virology Authors: Cui L, Mahesutihan M, Zheng W, Meng L, Fan W, Li J, Ye X, Liu W, Sun L Tags: Virology Source Type: research

Inhibition of influenza A virus matrix and nonstructural gene expression using RNA interference.
Abstract Influenza antiviral drugs that use protein inhibitors can lose their efficacy as resistant strains emerge. As an alternative strategy, we investigated the use of small interfering RNA molecules (siRNAs) by characterizing three siRNAs (M747, M776 and M832) targeting the influenza matrix 2 gene and three (NS570, NS595 and NS615) targeting the nonstructural protein 1 and 2 genes. We also re-examined two previously reported siRNAs, M331 and M950, which target the matrix 1 and 2 genes. Treatment with M331-, M776-, M832-, and M950-siRNAs attenuated influenza titer. M776-siRNA treated cells had 29.8% less infect...
Source: Virology - July 26, 2016 Category: Virology Authors: McMillen CM, Beezhold DH, Blachere FM, Othumpangat S, Kashon ML, Noti JD Tags: Virology Source Type: research

Hemagglutinin of influenza A virus binds specifically to cell surface nucleolin and plays a role in virus internalization.
Abstract The hemagglutinin (HA) protein of influenza A virus initiates cell entry by binding to sialic acids on target cells. In the current study, we demonstrated that in addition to sialic acids, influenza A/Puerto Rico/8/34 H1N1 (PR8) virus HA specifically binds to cell surface nucleolin (NCL). The interaction between HA and NCL was initially revealed with virus overlay protein binding assay (VOPBA) and subsequently verified with co-immunoprecipitation. Importantly, inhibiting cell surface NCL with NCL antibody, blocking PR8 viruses with purified NCL protein, or depleting endogenous NCL with siRNA all substanti...
Source: Virology - April 12, 2016 Category: Virology Authors: Chan CM, Chu H, Zhang AJ, Leung LH, Sze KH, Kao RY, Chik KK, To KK, Chan JF, Chen H, Jin DY, Liu L, Yuen KY Tags: Virology Source Type: research

ICAM-1 regulates the survival of influenza virus in lung epithelial cells during the early stages of infection.
Abstract Intercellular cell adhesion molecule-1 (ICAM-1) is an inducible cell surface glycoprotein that is expressed on many cell types. Influenza virus infection enhanced ICAM-1 expression and messenger RNA levels. Human bronchial epithelial cells (HBEpC) and nasal epithelial cells, on exposure to different strains of influenza virus (H1N1, H3N2, and H9N1) showed significant increase in ICAM-1 gene expression (p<0.001) along with the ICAM-1 protein levels (surface and secreted). Depleting ICAM-1 in HBEpC with ICAM-1 siRNA and subsequently infecting with H1N1 showed increased viral copy numbers. Influenza virus...
Source: Virology - October 22, 2015 Category: Virology Authors: Othumpangat S, Noti JD, McMillen CM, Beezhold DH Tags: Virology Source Type: research

RIG-I and TLR3 are both required for maximum interferon induction by influenza virus in human lung alveolar epithelial cells.
Abstract Pattern recognition receptors, such as retinoic acid-inducible protein I (RIG-I), Toll-like receptors 3 and 7 (TLR3 and 7), and nucleotide-binding oligomerization domain containing protein 2 (NOD2), play important roles in the recognition of influenza A virus (IAV), but their role in interferon (IFN) induction is still unclear, particularly in human lung. We investigated IFN induction by IAV in the A549 cell line as well as in primary human alveolar epithelial cells (AEC). TLR3/7, NOD2, RIG-I, and IFN expression levels were measured by qRT-PCR and ELISA in cells infected with IAV PR8. We found that TLR7 a...
Source: Virology - April 10, 2015 Category: Virology Authors: Wu W, Zhang W, Duggan ES, Booth JL, Zou MH, Metcalf JP Tags: Virology Source Type: research

hnRNP A2/B1 interacts with influenza A viral protein NS1 and inhibits virus replication potentially through suppressing NS1 RNA/protein levels and NS1 mRNA nuclear export.
In this study, we identified heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) as an interacting partner of NS1 proteins by a proteomic method. Knockdown of hnRNP A2/B1 by small interfering RNA (siRNA) resulted in higher levels of NS vRNA, NS1 mRNA, and NS1 protein in the virus-infected cells. In addition, we demonstrated that hnRNP A2/B1 proteins are associated with NS1 and NS2 mRNAs and that knockdown of hnRNP A2/B1 promotes transport of NS1 mRNA from the nucleus to the cytoplasm in the infected cells. Lastly, we showed that knockdown of hnRNP A2/B1 leads to enhanced virus replication. Our results suggest that ...
Source: Virology - January 18, 2014 Category: Virology Authors: Wang Y, Zhou J, Du Y Tags: Virology Source Type: research