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Developing a tool to shoot genes by a man-made air pressure

BACKGROUND: Biolistic systems are used to shoot exogenous DNA, RNA, protein, and other macromolecules to transfer them into cells for genetic transformation, genome editing, and drug delivery. Such systems are especially useful for plants and other organisms that are incompatible with other macromol...

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Autores principales: Tsugama, Daisuke, Takano, Tetsuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7486349/
https://www.ncbi.nlm.nih.gov/pubmed/32915413
http://dx.doi.org/10.1186/s43141-020-00067-1
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author Tsugama, Daisuke
Takano, Tetsuo
author_facet Tsugama, Daisuke
Takano, Tetsuo
author_sort Tsugama, Daisuke
collection PubMed
description BACKGROUND: Biolistic systems are used to shoot exogenous DNA, RNA, protein, and other macromolecules to transfer them into cells for genetic transformation, genome editing, and drug delivery. Such systems are especially useful for plants and other organisms that are incompatible with other macromolecule delivery methods. Commercially available, conventional biolistic systems consist of a shooting device (or “gun”) and a cylinder bottle for high-pressure helium gas. These cost a lot for installation and have low portability. RESULTS: We assembled an inexpensive air duster gun and a hand pump into a portable tool to shoot genes by a man-made air pressure (TSGAMAP). TSGAMAP allows to shoot DNA-coated gold particles with the 3-MPa maximum air pressure. When DNA with a fluorescent protein gene, GFP, was shot by TSGAMAP into leaf epidermal cells of onion, leaf lettuce, and Chinese cabbage, for all of these species, some cells in all became to exhibit GFP signals. When GFP was shot with another fluorescent protein gene, mCherry, into Chinese cabbage cells, both GFP and mCherry signals were detected in some cells. When a transcription factor gene AoAMS was fused with GFP and shot into Chinese cabbage cells, nuclear-localized GFP signals were detected in some cells. These results suggest that TSGAMAP can be used for protein coexpression and protein subcellular localization analyses. CONCLUSIONS: TSGAMAP is a cost-saving and portable tool to shoot DNA and other microparticles into cells. This can expand the use of biolistics in research and education.
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spelling pubmed-74863492020-09-21 Developing a tool to shoot genes by a man-made air pressure Tsugama, Daisuke Takano, Tetsuo J Genet Eng Biotechnol Research BACKGROUND: Biolistic systems are used to shoot exogenous DNA, RNA, protein, and other macromolecules to transfer them into cells for genetic transformation, genome editing, and drug delivery. Such systems are especially useful for plants and other organisms that are incompatible with other macromolecule delivery methods. Commercially available, conventional biolistic systems consist of a shooting device (or “gun”) and a cylinder bottle for high-pressure helium gas. These cost a lot for installation and have low portability. RESULTS: We assembled an inexpensive air duster gun and a hand pump into a portable tool to shoot genes by a man-made air pressure (TSGAMAP). TSGAMAP allows to shoot DNA-coated gold particles with the 3-MPa maximum air pressure. When DNA with a fluorescent protein gene, GFP, was shot by TSGAMAP into leaf epidermal cells of onion, leaf lettuce, and Chinese cabbage, for all of these species, some cells in all became to exhibit GFP signals. When GFP was shot with another fluorescent protein gene, mCherry, into Chinese cabbage cells, both GFP and mCherry signals were detected in some cells. When a transcription factor gene AoAMS was fused with GFP and shot into Chinese cabbage cells, nuclear-localized GFP signals were detected in some cells. These results suggest that TSGAMAP can be used for protein coexpression and protein subcellular localization analyses. CONCLUSIONS: TSGAMAP is a cost-saving and portable tool to shoot DNA and other microparticles into cells. This can expand the use of biolistics in research and education. Springer Berlin Heidelberg 2020-09-11 /pmc/articles/PMC7486349/ /pubmed/32915413 http://dx.doi.org/10.1186/s43141-020-00067-1 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Research
Tsugama, Daisuke
Takano, Tetsuo
Developing a tool to shoot genes by a man-made air pressure
title Developing a tool to shoot genes by a man-made air pressure
title_full Developing a tool to shoot genes by a man-made air pressure
title_fullStr Developing a tool to shoot genes by a man-made air pressure
title_full_unstemmed Developing a tool to shoot genes by a man-made air pressure
title_short Developing a tool to shoot genes by a man-made air pressure
title_sort developing a tool to shoot genes by a man-made air pressure
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7486349/
https://www.ncbi.nlm.nih.gov/pubmed/32915413
http://dx.doi.org/10.1186/s43141-020-00067-1
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