Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus
RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causa...
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PeerJ Inc.
2021-06-01
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| Online Access: | https://peerj.com/articles/11227.pdf |
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| author | Anabella Currá Marco Cacciabue María José Gravisaco Sebastián Asurmendi Oscar Taboga María I. Gismondi |
| author_facet | Anabella Currá Marco Cacciabue María José Gravisaco Sebastián Asurmendi Oscar Taboga María I. Gismondi |
| author_sort | Anabella Currá |
| collection | DOAJ |
| description | RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causative agent of a devastating animal disease with high economic impact. The aim of this work was to assess the antiviral activity of different small RNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D). Three target sequences were predicted within 3D considering RNA accessibility as a major criterion. The silencing efficacy of short-hairpin RNAs (shRNAs) and artificial microRNAs (amiRNAs) targeting the selected sequences was confirmed in fluorescent reporter assays. Furthermore, BHK-21 cells transiently expressing shRNAs or amiRNAs proved 70 to >95% inhibition of FMDV growth. Interestingly, dual expression of amiRNAs did not improve FMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs were established and characterized in terms of antiviral activity against FMDV. As expected, viral replication in these cell lines was delayed. These results show that the target RNA-accessibility-guided approach for RNAi design rendered efficient amiRNAs that constrain FMDV replication. The application of amiRNAs to complement FMDV vaccination in specific epidemiological scenarios shall be explored further. |
| first_indexed | 2024-03-09T06:32:36Z |
| format | Article |
| id | doaj.art-fbaa6adcbcd6404b8339913ea9b87134 |
| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:32:36Z |
| publishDate | 2021-06-01 |
| publisher | PeerJ Inc. |
| record_format | Article |
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| spelling | doaj.art-fbaa6adcbcd6404b8339913ea9b871342023-12-03T11:03:58ZengPeerJ Inc.PeerJ2167-83592021-06-019e1122710.7717/peerj.11227Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virusAnabella Currá0Marco Cacciabue1María José Gravisaco2Sebastián Asurmendi3Oscar Taboga4María I. Gismondi5Instituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaInstituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaInstituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaInstituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaInstituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaInstituto de Agrobiotecnología y Biología Molecular (IABiMo), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Hurlingham, Buenos Aires, ArgentinaRNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causative agent of a devastating animal disease with high economic impact. The aim of this work was to assess the antiviral activity of different small RNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D). Three target sequences were predicted within 3D considering RNA accessibility as a major criterion. The silencing efficacy of short-hairpin RNAs (shRNAs) and artificial microRNAs (amiRNAs) targeting the selected sequences was confirmed in fluorescent reporter assays. Furthermore, BHK-21 cells transiently expressing shRNAs or amiRNAs proved 70 to >95% inhibition of FMDV growth. Interestingly, dual expression of amiRNAs did not improve FMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs were established and characterized in terms of antiviral activity against FMDV. As expected, viral replication in these cell lines was delayed. These results show that the target RNA-accessibility-guided approach for RNAi design rendered efficient amiRNAs that constrain FMDV replication. The application of amiRNAs to complement FMDV vaccination in specific epidemiological scenarios shall be explored further.https://peerj.com/articles/11227.pdfFMDVAntiviralsRNA interferenceTarget accessibility |
| spellingShingle | Anabella Currá Marco Cacciabue María José Gravisaco Sebastián Asurmendi Oscar Taboga María I. Gismondi Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus PeerJ FMDV Antivirals RNA interference Target accessibility |
| title | Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus |
| title_full | Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus |
| title_fullStr | Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus |
| title_full_unstemmed | Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus |
| title_short | Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus |
| title_sort | antiviral efficacy of short hairpin rnas and artificial micrornas targeting foot and mouth disease virus |
| topic | FMDV Antivirals RNA interference Target accessibility |
| url | https://peerj.com/articles/11227.pdf |
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