Abstract
The knowledge that small RNAs can affect gene expression has had a tremendous impact on basic and applied research, and gene silencing is currently one of the most promising new approaches for disease therapy. However, RNAs cannot easily penetrate cell membranes, therefore RNA delivery become one of the major challenges for gene silencing technology. In the current paper we discuss a general approach for converting siRNA molecules into a dense siRNA nanoparticles using environmentally friendly sonochemical method. The RNA nanoparticulation enhance its gene-silencing activity in vascular bovine endothelial as well as in cancer 293T/GFP-Puro cell lines without causing any toxic effect. We show that ultrasonic waves do not lead to RNA degradation or any changes in its chemical structure. Moreover, sonochemically produced siRNA nanoparticles have been shown to be resistant to a variety of environmental stresses including pH levels, enzymes and temperatures, hence solving problem of the short half-life of the RNA molecules. As the siRNA nanoparticles are biocompatibile and biodegradabile, and their RNA release properties may be controlled within limits, sonochemical formation of siRNA nanoparticles represent a new promising approach for generation of functional bionano materials.
Highlights
RNA, has recently emerged as an important nanotechnology platform due to its extraordinary diversity in structure and function [1,2]
Introduction of Small interfering RNA (siRNA) nanoparticles into the cells and gene silencing activity assay Vascular endothelial cells were prepared from segments of the descending bovine aorta obtained from a local slaughterhouse (Kibbutz Lahav, Israel)
We further investigated the dependence of the morphology and size of the siRNA nanoparticles formed upon different exposure times to ultrasonic waves by high resolution scanning electron microscopy (HRSEM) and scanning transmission electron microscopy (STEM) (Figure 1a)
Summary
RNA, has recently emerged as an important nanotechnology platform due to its extraordinary diversity in structure and function [1,2]. The sonochemical siRNA nanoparticulation stabilize siRNA molecules In this manner silencing activity of siRNA nanoparticles achieved through its dissociation to individual RNA molecules inside the cells. We probed the gene-silencing properties of nanoparticulate siRNA for endothelial bovine [18,19] and 293T/ GFP-Puro [20] cell cultures. Altered structure and function of endothelial cells might contribute significantly to the pathogenesis of disorders or the arterial wall. This is a not easy task to lead to penetration of siRNA to normal, not cancer cells. SiRNA nanoparticles into the cells and gene silencing activity assay Vascular endothelial cells were prepared from segments of the descending bovine aorta obtained from a local slaughterhouse (Kibbutz Lahav, Israel). The internalization of the nanoparticles inside the cells was visualized using confocal microscopy
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