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In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes

Dengue virus is a ssRNA+ flavivirus, which produces the dengue disease in humans. Currently, no specific treatment exists. siRNAs regulate gene expression and have been used systematically to silence viral genomes; however, they require controlled release. Liposomes show favorable results encapsulat...

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Autores principales: Rodriguez-Salazar, Carlos Andrés, Recalde-Reyes, Delia Piedad, Bedoya, Juan Pablo, Padilla-Sanabria, Leonardo, Castaño-Osorio, Jhon Carlos, Giraldo, Maria Isabel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875542/
https://www.ncbi.nlm.nih.gov/pubmed/35215929
http://dx.doi.org/10.3390/v14020339
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author Rodriguez-Salazar, Carlos Andrés
Recalde-Reyes, Delia Piedad
Bedoya, Juan Pablo
Padilla-Sanabria, Leonardo
Castaño-Osorio, Jhon Carlos
Giraldo, Maria Isabel
author_facet Rodriguez-Salazar, Carlos Andrés
Recalde-Reyes, Delia Piedad
Bedoya, Juan Pablo
Padilla-Sanabria, Leonardo
Castaño-Osorio, Jhon Carlos
Giraldo, Maria Isabel
author_sort Rodriguez-Salazar, Carlos Andrés
collection PubMed
description Dengue virus is a ssRNA+ flavivirus, which produces the dengue disease in humans. Currently, no specific treatment exists. siRNAs regulate gene expression and have been used systematically to silence viral genomes; however, they require controlled release. Liposomes show favorable results encapsulating siRNA for gene silencing. The objective herein was to design and evaluate in vitro siRNAs bound to liposomes that inhibit DENV replication. siRNAs were designed against DENV1–4 from conserved regions using siDirect2.0 and Web-BLOCK-iT™ RNAiDesigner; the initial in vitro evaluation was carried out through transfection into HepG2 cells. siRNA with silencing capacity was encapsulated in liposomes composed of D-Lin-MC3-DMA, DSPC, Chol. Cytotoxicity, hemolysis, pro-inflammatory cytokine release and antiviral activity were evaluated using plaque assay and RT-qPCR. A working concentration of siRNA was established at 40 nM. siRNA1, siRNA2, siRNA3.1, and siRNA4 were encapsulated in liposomes, and their siRNA delivery through liposomes led to a statistically significant decrease in viral titers, yielded no cytotoxicity or hemolysis and did not stimulate release of pro-inflammatory cytokines. Finally, liposomes were designed with siRNA against DENV, which proved to be safe in vitro.
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spelling pubmed-88755422022-02-26 In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes Rodriguez-Salazar, Carlos Andrés Recalde-Reyes, Delia Piedad Bedoya, Juan Pablo Padilla-Sanabria, Leonardo Castaño-Osorio, Jhon Carlos Giraldo, Maria Isabel Viruses Article Dengue virus is a ssRNA+ flavivirus, which produces the dengue disease in humans. Currently, no specific treatment exists. siRNAs regulate gene expression and have been used systematically to silence viral genomes; however, they require controlled release. Liposomes show favorable results encapsulating siRNA for gene silencing. The objective herein was to design and evaluate in vitro siRNAs bound to liposomes that inhibit DENV replication. siRNAs were designed against DENV1–4 from conserved regions using siDirect2.0 and Web-BLOCK-iT™ RNAiDesigner; the initial in vitro evaluation was carried out through transfection into HepG2 cells. siRNA with silencing capacity was encapsulated in liposomes composed of D-Lin-MC3-DMA, DSPC, Chol. Cytotoxicity, hemolysis, pro-inflammatory cytokine release and antiviral activity were evaluated using plaque assay and RT-qPCR. A working concentration of siRNA was established at 40 nM. siRNA1, siRNA2, siRNA3.1, and siRNA4 were encapsulated in liposomes, and their siRNA delivery through liposomes led to a statistically significant decrease in viral titers, yielded no cytotoxicity or hemolysis and did not stimulate release of pro-inflammatory cytokines. Finally, liposomes were designed with siRNA against DENV, which proved to be safe in vitro. MDPI 2022-02-07 /pmc/articles/PMC8875542/ /pubmed/35215929 http://dx.doi.org/10.3390/v14020339 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rodriguez-Salazar, Carlos Andrés
Recalde-Reyes, Delia Piedad
Bedoya, Juan Pablo
Padilla-Sanabria, Leonardo
Castaño-Osorio, Jhon Carlos
Giraldo, Maria Isabel
In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title_full In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title_fullStr In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title_full_unstemmed In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title_short In Vitro Inhibition of Replication of Dengue Virus Serotypes 1–4 by siRNAs Bound to Non-Toxic Liposomes
title_sort in vitro inhibition of replication of dengue virus serotypes 1–4 by sirnas bound to non-toxic liposomes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875542/
https://www.ncbi.nlm.nih.gov/pubmed/35215929
http://dx.doi.org/10.3390/v14020339
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