Abstract
BackgroundSmall interfering RNA (siRNA) has attracted attention in the field of nucleic acid medicine as a RNA interference (RNAi) application that leads to gene silencing due to specific messenger RNA (mRNA) destruction. However, since siRNA is unstable in blood and unable to cross the cell membrane, encapsulation of siRNA into a carrier is required.ResultsIn this study, we used a carrier that combined ZHER2-displaying bio-nanocapsule (derived from hepatitis B virus surface antigen) and liposomes in a complex in order to investigate the feasibility of effective and target-cell-specific RNAi applications. As a result, by observing RNAi only in HER2-expressing breast cancer cells, using our proposed methodology, we successfully demonstrated target-cell-specific delivery and effective function expression of siRNA.ConclusionsThese findings show that, in the field of nucleic acid medicine, ZHER2-BNC/LP can be a useful carrier for siRNA delivery, and could also become a useful tool for gene silencing and to accomplish protein knock-down.
Highlights
Small interfering RNA has attracted attention in the field of nucleic acid medicine as a RNA interference (RNAi) application that leads to gene silencing due to specific messenger RNA destruction
Yeast cells transformed with pGLDsLd50-ZHER2 or pGLDsLd50-ZWT by the spheroplast method were cultured and disrupted with glass beads, the crude extract was precipitated with polyethylene glycol (PEG) 6000 and subjected to cesium chloride (CsCl) isopycnic ultracentrifugation and sucrose density gradient ultracentrifugation, and the purified BNCs were obtained after freeze-drying in the presence of 5% sucrose
RNAiMAX, LPs and ZHER2-BNC/LP were tested to deliver silencer Green fluorescent protein (GFP) Small interfering RNA (siRNA) which can interfere with the expression of GFP
Summary
Small interfering RNA (siRNA) has attracted attention in the field of nucleic acid medicine as a RNA interference (RNAi) application that leads to gene silencing due to specific messenger RNA (mRNA) destruction. RNA interference (RNAi) is expected to become a new approach in treating a variety of diseases, such as virus infection, cancer and neurodegenerative diseases, owing to specific and effective gene silencing [1,2]. The mechanism of RNAi involves double-stranded RNA injected into cells that are first cut into short RNA (small interfering RNA (siRNA)), 21–23 bp long, using ribonuclease (RNase) III enzyme that is referred to as the Dicer. The duplex siRNA forms a RNA-induced silencing complex (RISC), which contains an endonuclease and an Argonaute protein. The siRNA duplex is dissociated into unwound single-stranded RNA using an ATP-dependent helicase; the RISC with an antisense strand against target messenger RNA (mRNA) leads to RNA. The BNC has been studied as a carrier for the delivery of drugs and genes [9]
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