期刊
VIRUSES-BASEL
卷 14, 期 2, 页码 -出版社
MDPI
DOI: 10.3390/v14020339
关键词
dengue virus; liposomes; siRNA
类别
资金
- Corporacion Universitaria Empresarial Alexander von Humboldt [948]
- National Institutes of Health/Office of the Director (OD)/National Institute of Allergy and Infectious Diseases (NIAID) [K12HD052023]
- Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
This study designed and evaluated the combination of siRNA and liposomes to inhibit dengue virus replication. siRNA delivered through liposomes significantly reduced viral titers, without cytotoxicity, hemolysis, or stimulation of pro-inflammatory cytokine release. These results suggest that liposomal delivery of siRNA is a safe and effective method to inhibit dengue virus.
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 (TM) 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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据