SiRNA against the G gene of human metapneumovirus

Faith Preston, Claire Straub, Suresh Mahalingam, Kirsten Spann

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background Human metapneumovirus (hMPV) is a significant viral respiratory pathogen of infants and children, the elderly and immunocompromised individuals. Disease associated with hMPV infection resembles that of human respiratory syncytial virus (RSV) and includes bronchiolitis and pneumonia. The glycosylated G attachment protein of hMPV is required for viral entry in vivo and has also been identified as an inhibitor of innate immune responses. Findings We designed and validated two siRNA molecules against the G gene using A549 cells and demonstrated consistent 88-92% knock-down for one siRNA molecule, which was used in subsequent experiments. Significant reduction of G mRNA in A549 cells infected with hMPV did not result in a reduction in viral growth, nor did it significantly increase the production of type I interferon (α/β) in response to infection. However, there was a moderate increase in IFN-β mRNA expression in response to infection in siG-transfected cells compared to untransfected and si-mismatch-transfected cells. Expression of G by recombinant adenovirus did not affect type I IFN expression. Conclusion G has been previously described as a type I interferon antagonist, although our findings suggest it may not be a significant antagonist.
Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalVirology Journal
Volume9
DOIs
Publication statusPublished - 2012
Externally publishedYes

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Metapneumovirus
Interferon Type I
Small Interfering RNA
Genes
Infection
Human respiratory syncytial virus
Messenger RNA
Bronchiolitis
Innate Immunity
Adenoviridae
Pneumonia
Growth
A549 Cells

Cite this

Preston, F., Straub, C., Mahalingam, S., & Spann, K. (2012). SiRNA against the G gene of human metapneumovirus. Virology Journal, 9, 1-5. https://doi.org/10.1186/1743-422X-9-105
Preston, Faith ; Straub, Claire ; Mahalingam, Suresh ; Spann, Kirsten. / SiRNA against the G gene of human metapneumovirus. In: Virology Journal. 2012 ; Vol. 9. pp. 1-5.
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Preston, F, Straub, C, Mahalingam, S & Spann, K 2012, 'SiRNA against the G gene of human metapneumovirus', Virology Journal, vol. 9, pp. 1-5. https://doi.org/10.1186/1743-422X-9-105

SiRNA against the G gene of human metapneumovirus. / Preston, Faith; Straub, Claire; Mahalingam, Suresh; Spann, Kirsten.

In: Virology Journal, Vol. 9, 2012, p. 1-5.

Research output: Contribution to journalArticle

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AU - Preston, Faith

AU - Straub, Claire

AU - Mahalingam, Suresh

AU - Spann, Kirsten

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N2 - Background Human metapneumovirus (hMPV) is a significant viral respiratory pathogen of infants and children, the elderly and immunocompromised individuals. Disease associated with hMPV infection resembles that of human respiratory syncytial virus (RSV) and includes bronchiolitis and pneumonia. The glycosylated G attachment protein of hMPV is required for viral entry in vivo and has also been identified as an inhibitor of innate immune responses. Findings We designed and validated two siRNA molecules against the G gene using A549 cells and demonstrated consistent 88-92% knock-down for one siRNA molecule, which was used in subsequent experiments. Significant reduction of G mRNA in A549 cells infected with hMPV did not result in a reduction in viral growth, nor did it significantly increase the production of type I interferon (α/β) in response to infection. However, there was a moderate increase in IFN-β mRNA expression in response to infection in siG-transfected cells compared to untransfected and si-mismatch-transfected cells. Expression of G by recombinant adenovirus did not affect type I IFN expression. Conclusion G has been previously described as a type I interferon antagonist, although our findings suggest it may not be a significant antagonist.

AB - Background Human metapneumovirus (hMPV) is a significant viral respiratory pathogen of infants and children, the elderly and immunocompromised individuals. Disease associated with hMPV infection resembles that of human respiratory syncytial virus (RSV) and includes bronchiolitis and pneumonia. The glycosylated G attachment protein of hMPV is required for viral entry in vivo and has also been identified as an inhibitor of innate immune responses. Findings We designed and validated two siRNA molecules against the G gene using A549 cells and demonstrated consistent 88-92% knock-down for one siRNA molecule, which was used in subsequent experiments. Significant reduction of G mRNA in A549 cells infected with hMPV did not result in a reduction in viral growth, nor did it significantly increase the production of type I interferon (α/β) in response to infection. However, there was a moderate increase in IFN-β mRNA expression in response to infection in siG-transfected cells compared to untransfected and si-mismatch-transfected cells. Expression of G by recombinant adenovirus did not affect type I IFN expression. Conclusion G has been previously described as a type I interferon antagonist, although our findings suggest it may not be a significant antagonist.

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