Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery
Wen Zhao, Yifan Zhang, Xueyun Jiang, Chunying Cui School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China Abstract: Small interfering RNA (siRNA) delivery is a prospective method in gene therapy, but it has application limitations such as negative charge,...
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doaj-07ce8d8e171d4f95876a94cace895fc12020-11-24T22:31:48ZengDove Medical PressDrug Design, Development and Therapy1177-88812016-11-01Volume 103851386530211Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene deliveryZhao WZhang YFJiang XYCui CYWen Zhao, Yifan Zhang, Xueyun Jiang, Chunying Cui School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China Abstract: Small interfering RNA (siRNA) delivery is a prospective method in gene therapy, but it has application limitations such as negative charge, water solubility and high molecular weight. In this study, a safe and efficient nano-vector, CRS, was designed and synthesized to facilitate siRNA delivery. Physical and chemical properties of VEGF-siRNA/CRS were characterized by methods including scanning electron microscopy (SEM), transmission electron microscopy, zeta potential (ζ) measurement, drug-releasing rate measurement, gel electrophoresis and confocal microscopy. The biological activities were evaluated using cell viability assay, gene-silencing efficacy assay in vitro, real-time polymerase chain reaction, enzyme-linked immunosorbent assay (ELISA) and antitumor tests in vivo. The mean nanoparticle size of VEGF-siRNA/CRS was 121.4±0.3 nm with positive ζ potential of 7.69±4.47 mV. The release rate of VEGF-siRNA from VEGF-siRNA/CRS was 82.50% sustained for 48 h in Tris-ethylenediaminetetraacetic acid buffer (pH 8.0). Real-time polymerase chain reaction was used to analyze the efficiency of the transfection, and the result showed that VEGF mRNA expression had been knocked down by 82.36%. The expression of VEGF protein was also recorded to be downregulated to 14.83% using ELISA. The results of cytotoxicity measured by Cell Counting Kit-8 assay showed that VEGF-siRNA/CRS had significant inhibitory effect on HeLa cells. The results of antitumor assays indicated that VEGF-siRNA/CRS exhibited tumor cell growth inhibition in vivo. The results demonstrated that VEGF-siRNA could be delivered and transported by the designed carrier, while siRNA could be released constantly and led to an increasing gene-silencing effect against VEGF gene. In conclusion, VEGF-siRNA/CRS is a promising carrier for siRNA delivery, and further studies are warranted. Keywords: CRS, siRNA delivery, VEGF, tumor therapy, antitumor, biophysical characteristics, gene silencing, protein expressionhttps://www.dovepress.com/design-synthesis-and-evaluation-of-vegf-sirnacrs-as-a-novel-vector-for-peer-reviewed-article-DDDTCRSsiRNA deliveryVEGFtumor therapy (anti-tumor)biophysical characteristicsgene silencingprotein expression. |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zhao W Zhang YF Jiang XY Cui CY |
spellingShingle |
Zhao W Zhang YF Jiang XY Cui CY Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery Drug Design, Development and Therapy CRS siRNA delivery VEGF tumor therapy (anti-tumor) biophysical characteristics gene silencing protein expression. |
author_facet |
Zhao W Zhang YF Jiang XY Cui CY |
author_sort |
Zhao W |
title |
Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery |
title_short |
Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery |
title_full |
Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery |
title_fullStr |
Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery |
title_full_unstemmed |
Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery |
title_sort |
design, synthesis and evaluation of vegf-sirna/crs as a novel vector for gene delivery |
publisher |
Dove Medical Press |
series |
Drug Design, Development and Therapy |
issn |
1177-8881 |
publishDate |
2016-11-01 |
description |
Wen Zhao, Yifan Zhang, Xueyun Jiang, Chunying Cui School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China Abstract: Small interfering RNA (siRNA) delivery is a prospective method in gene therapy, but it has application limitations such as negative charge, water solubility and high molecular weight. In this study, a safe and efficient nano-vector, CRS, was designed and synthesized to facilitate siRNA delivery. Physical and chemical properties of VEGF-siRNA/CRS were characterized by methods including scanning electron microscopy (SEM), transmission electron microscopy, zeta potential (ζ) measurement, drug-releasing rate measurement, gel electrophoresis and confocal microscopy. The biological activities were evaluated using cell viability assay, gene-silencing efficacy assay in vitro, real-time polymerase chain reaction, enzyme-linked immunosorbent assay (ELISA) and antitumor tests in vivo. The mean nanoparticle size of VEGF-siRNA/CRS was 121.4±0.3 nm with positive ζ potential of 7.69±4.47 mV. The release rate of VEGF-siRNA from VEGF-siRNA/CRS was 82.50% sustained for 48 h in Tris-ethylenediaminetetraacetic acid buffer (pH 8.0). Real-time polymerase chain reaction was used to analyze the efficiency of the transfection, and the result showed that VEGF mRNA expression had been knocked down by 82.36%. The expression of VEGF protein was also recorded to be downregulated to 14.83% using ELISA. The results of cytotoxicity measured by Cell Counting Kit-8 assay showed that VEGF-siRNA/CRS had significant inhibitory effect on HeLa cells. The results of antitumor assays indicated that VEGF-siRNA/CRS exhibited tumor cell growth inhibition in vivo. The results demonstrated that VEGF-siRNA could be delivered and transported by the designed carrier, while siRNA could be released constantly and led to an increasing gene-silencing effect against VEGF gene. In conclusion, VEGF-siRNA/CRS is a promising carrier for siRNA delivery, and further studies are warranted. Keywords: CRS, siRNA delivery, VEGF, tumor therapy, antitumor, biophysical characteristics, gene silencing, protein expression |
topic |
CRS siRNA delivery VEGF tumor therapy (anti-tumor) biophysical characteristics gene silencing protein expression. |
url |
https://www.dovepress.com/design-synthesis-and-evaluation-of-vegf-sirnacrs-as-a-novel-vector-for-peer-reviewed-article-DDDT |
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