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Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance

The use of syn-tasiRNAs has been proposed as an RNA interference technique alternative to those previously described: hairpin based, virus induced gene silencing or artificial miRNAs. In this study we engineered the TAS1c locus to impair Plum pox virus (PPV) infection by replacing the five native si...

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Autores principales: Zhao, Mingmin, San León, David, Mesel, Frida, García, Juan Antonio, Simón-Mateo, Carmen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4492489/
https://www.ncbi.nlm.nih.gov/pubmed/26147769
http://dx.doi.org/10.1371/journal.pone.0132281
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author Zhao, Mingmin
San León, David
Mesel, Frida
García, Juan Antonio
Simón-Mateo, Carmen
author_facet Zhao, Mingmin
San León, David
Mesel, Frida
García, Juan Antonio
Simón-Mateo, Carmen
author_sort Zhao, Mingmin
collection PubMed
description The use of syn-tasiRNAs has been proposed as an RNA interference technique alternative to those previously described: hairpin based, virus induced gene silencing or artificial miRNAs. In this study we engineered the TAS1c locus to impair Plum pox virus (PPV) infection by replacing the five native siRNAs with two 210-bp fragments from the CP and the 3´NCR regions of the PPV genome. Deep sequencing analysis of the small RNA species produced by both constructs in planta has shown that phased processing of the syn-tasiRNAs is construct-specific. While in syn-tasiR-CP construct the processing was as predicted 21-nt phased in register with miR173-guided cleavage, the processing of syn-tasiR-3NCR is far from what was expected. A 22-nt species from the miR173-guided cleavage was a guide of two series of phased small RNAs, one of them in an exact 21-nt register, and the other one in a mixed of 21-/22-nt frame. In addition, both constructs produced abundant PPV-derived small RNAs in the absence of miR173 as a consequence of a strong sense post-transcriptional gene silencing induction. The antiviral effect of both constructs was also evaluated in the presence or absence of miR173 and showed that the impairment of PPV infection was not significantly higher when miR173 was present. The results show that syn-tasiRNAs processing depends on construct-specific factors that should be further studied before the so-called MIGS (miRNA-induced gene silencing) technology can be used reliably.
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spelling pubmed-44924892015-07-15 Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance Zhao, Mingmin San León, David Mesel, Frida García, Juan Antonio Simón-Mateo, Carmen PLoS One Research Article The use of syn-tasiRNAs has been proposed as an RNA interference technique alternative to those previously described: hairpin based, virus induced gene silencing or artificial miRNAs. In this study we engineered the TAS1c locus to impair Plum pox virus (PPV) infection by replacing the five native siRNAs with two 210-bp fragments from the CP and the 3´NCR regions of the PPV genome. Deep sequencing analysis of the small RNA species produced by both constructs in planta has shown that phased processing of the syn-tasiRNAs is construct-specific. While in syn-tasiR-CP construct the processing was as predicted 21-nt phased in register with miR173-guided cleavage, the processing of syn-tasiR-3NCR is far from what was expected. A 22-nt species from the miR173-guided cleavage was a guide of two series of phased small RNAs, one of them in an exact 21-nt register, and the other one in a mixed of 21-/22-nt frame. In addition, both constructs produced abundant PPV-derived small RNAs in the absence of miR173 as a consequence of a strong sense post-transcriptional gene silencing induction. The antiviral effect of both constructs was also evaluated in the presence or absence of miR173 and showed that the impairment of PPV infection was not significantly higher when miR173 was present. The results show that syn-tasiRNAs processing depends on construct-specific factors that should be further studied before the so-called MIGS (miRNA-induced gene silencing) technology can be used reliably. Public Library of Science 2015-07-06 /pmc/articles/PMC4492489/ /pubmed/26147769 http://dx.doi.org/10.1371/journal.pone.0132281 Text en © 2015 Zhao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zhao, Mingmin
San León, David
Mesel, Frida
García, Juan Antonio
Simón-Mateo, Carmen
Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title_full Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title_fullStr Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title_full_unstemmed Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title_short Assorted Processing of Synthetic Trans-Acting siRNAs and Its Activity in Antiviral Resistance
title_sort assorted processing of synthetic trans-acting sirnas and its activity in antiviral resistance
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4492489/
https://www.ncbi.nlm.nih.gov/pubmed/26147769
http://dx.doi.org/10.1371/journal.pone.0132281
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