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...

Descripción completa

Detalles Bibliográficos
Autores principales: Curra, Anabella Paola, Cacciabue, Marco Polo Domingo, Gravisaco, María José, Asurmendi, Sebastian, Taboga, Oscar Alberto, Gismondi, Maria Ines
Formato: info:ar-repo/semantics/artículo
Lenguaje:Inglés
Publicado: Peer J Inc. 2024
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/18512
https://peerj.com/articles/11227/
https://doi.org/10.7717/peerj.11227
_version_ 1855037870620803072
author Curra, Anabella Paola
Cacciabue, Marco Polo Domingo
Gravisaco, María José
Asurmendi, Sebastian
Taboga, Oscar Alberto
Gismondi, Maria Ines
author_browse Asurmendi, Sebastian
Cacciabue, Marco Polo Domingo
Curra, Anabella Paola
Gismondi, Maria Ines
Gravisaco, María José
Taboga, Oscar Alberto
author_facet Curra, Anabella Paola
Cacciabue, Marco Polo Domingo
Gravisaco, María José
Asurmendi, Sebastian
Taboga, Oscar Alberto
Gismondi, Maria Ines
author_sort Curra, Anabella Paola
collection INTA Digital
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.
format info:ar-repo/semantics/artículo
id INTA18512
institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
language Inglés
publishDate 2024
publishDateRange 2024
publishDateSort 2024
publisher Peer J Inc.
publisherStr Peer J Inc.
record_format dspace
spelling INTA185122024-07-15T14:21:22Z Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot and-mouth disease virus Curra, Anabella Paola Cacciabue, Marco Polo Domingo Gravisaco, María José Asurmendi, Sebastian Taboga, Oscar Alberto Gismondi, Maria Ines Foot-and-mouth Disease Aphthovirus Antiviral Agents RNA Fiebre Aftosa Virus Fiebre Aftosa Viricida ARN 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. Instituto de Biotecnología Fil: Curra, Anabella Paola. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Curra, Anabella Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cacciabue, Marco Polo Domingo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Cacciabue, Marco Polo Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gravisaco, María José. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Gravisaco, María José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Asurmendi, Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Asurmendi, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gismondi, Maria Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Gismondi, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2024-07-15T14:06:11Z 2024-07-15T14:06:11Z 2021-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/18512 https://peerj.com/articles/11227/ 2167-8359 https://doi.org/10.7717/peerj.11227 eng info:eu-repograntAgreement/INTA/PNBIO/1131034/AR./Inmunología molecular y genómica funcional aplicadas a interacciones patógeno hospedador de interés pecuario. info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Peer J Inc. PeerJ 9 : e11227 (June 2021)
spellingShingle Foot-and-mouth Disease
Aphthovirus
Antiviral Agents
RNA
Fiebre Aftosa
Virus Fiebre Aftosa
Viricida
ARN
Curra, Anabella Paola
Cacciabue, Marco Polo Domingo
Gravisaco, María José
Asurmendi, Sebastian
Taboga, Oscar Alberto
Gismondi, Maria Ines
Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot and-mouth disease virus
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 Foot-and-mouth Disease
Aphthovirus
Antiviral Agents
RNA
Fiebre Aftosa
Virus Fiebre Aftosa
Viricida
ARN
url http://hdl.handle.net/20.500.12123/18512
https://peerj.com/articles/11227/
https://doi.org/10.7717/peerj.11227
work_keys_str_mv AT curraanabellapaola antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus
AT cacciabuemarcopolodomingo antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus
AT gravisacomariajose antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus
AT asurmendisebastian antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus
AT tabogaoscaralberto antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus
AT gismondimariaines antiviralefficacyofshorthairpinrnasandartificialmicrornastargetingfootandmouthdiseasevirus