Abstract
We have developed an in vivo transfection method for naked plasmid DNA (pDNA) and siRNA in mice by using a tissue suction device. The target tissue was suctioned by a device made of polydimethylsiloxane (PDMS) following the intravenous injection of naked pDNA or siRNA. Transfection of pDNA encoding luciferase was achieved by the suction of the kidney, liver, spleen, and heart, but not the duodenum, skeletal muscle, or stomach. Luciferase expression was specifically observed at the suctioned region of the tissue, and the highest luciferase expression was detected at the surface of the tissue (0.12±0.03 ng/mg protein in mice liver). Luciferase expression levels in the whole liver increased linearly with an increase in the number of times the liver was suctioned. Transfection of siRNA targeting glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene significantly suppressed the expression of GAPDH mRNA in the liver. Histological analysis shows that severe damage was not observed in the suctioned livers. Since the suction device can be mounted onto the head of the endoscope, this method is a minimally invasive. These results indicate that the in vivo transfection method developed in this study will be a viable approach for biological research and therapies using nucleic acids.
Highlights
In the post-genomic era, increased importance has been placed on the development of in vivo transfection techniques that can be used for biological research or gene therapy
It was shown that in vivo transfection of naked nucleic acids such as plasmid DNA (pDNA) and siRNA can be achieved by tissue suction
The present study demonstrates the first use of negative pressure to induce transfection
Summary
In the post-genomic era, increased importance has been placed on the development of in vivo transfection techniques that can be used for biological research or gene therapy. Transfection methods for naked nucleic acids, including plasmid DNA (pDNA) or siRNA, have many advantages, including convenient preparation, ease of handling, and lack of toxicity associated with the transfection agents. Our group found that direct pressure to the kidneys, spleen, and liver induces the transfection of naked nucleic acids, which we termed as tissue pressure-mediated transfection [8,9,10] This method has been used with naked pDNA, siRNA, and microRNA [8,9,10,11], and the miR-200 family of microRNAs introduced by renal pressure-mediated transfection ameliorated renal tubulointerstitial fibrosis in mice [11]. We previously reported that the secretion of pro-inflammatory cytokines was not observed under the experimental conditions for transfection and the degree of direct pressure applied to the target tissue is one of the key factors for controlling the expression levels of the transfected pDNA [9]
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