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In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device

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...

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Autores principales: Shimizu, Kazunori, Kawakami, Shigeru, Hayashi, Kouji, Kinoshita, Hideyuki, Kuwahara, Koichiro, Nakao, Kazuwa, Hashida, Mitsuru, Konishi, Satoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3402481/
https://www.ncbi.nlm.nih.gov/pubmed/22844458
http://dx.doi.org/10.1371/journal.pone.0041319
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author Shimizu, Kazunori
Kawakami, Shigeru
Hayashi, Kouji
Kinoshita, Hideyuki
Kuwahara, Koichiro
Nakao, Kazuwa
Hashida, Mitsuru
Konishi, Satoshi
author_facet Shimizu, Kazunori
Kawakami, Shigeru
Hayashi, Kouji
Kinoshita, Hideyuki
Kuwahara, Koichiro
Nakao, Kazuwa
Hashida, Mitsuru
Konishi, Satoshi
author_sort Shimizu, Kazunori
collection PubMed
description 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.
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spelling pubmed-34024812012-07-27 In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device Shimizu, Kazunori Kawakami, Shigeru Hayashi, Kouji Kinoshita, Hideyuki Kuwahara, Koichiro Nakao, Kazuwa Hashida, Mitsuru Konishi, Satoshi PLoS One Research Article 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. Public Library of Science 2012-07-23 /pmc/articles/PMC3402481/ /pubmed/22844458 http://dx.doi.org/10.1371/journal.pone.0041319 Text en Shimizu et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Shimizu, Kazunori
Kawakami, Shigeru
Hayashi, Kouji
Kinoshita, Hideyuki
Kuwahara, Koichiro
Nakao, Kazuwa
Hashida, Mitsuru
Konishi, Satoshi
In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title_full In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title_fullStr In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title_full_unstemmed In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title_short In vivo Site-Specific Transfection of Naked Plasmid DNA and siRNAs in Mice by Using a Tissue Suction Device
title_sort in vivo site-specific transfection of naked plasmid dna and sirnas in mice by using a tissue suction device
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3402481/
https://www.ncbi.nlm.nih.gov/pubmed/22844458
http://dx.doi.org/10.1371/journal.pone.0041319
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