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Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants

The capacity to introduce exogenous proteins and express (or down-regulate) specific genes in plants provides a powerful tool for fundamental research as well as new applications in the field of plant biotechnology. Viable methods that currently exist for protein or gene transfer into plant cells, n...

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Autores principales: Chuah, Jo-Ann, Horii, Yoko, Numata, Keiji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226412/
https://www.ncbi.nlm.nih.gov/pubmed/28060264
http://dx.doi.org/10.3791/54972
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author Chuah, Jo-Ann
Horii, Yoko
Numata, Keiji
author_facet Chuah, Jo-Ann
Horii, Yoko
Numata, Keiji
author_sort Chuah, Jo-Ann
collection PubMed
description The capacity to introduce exogenous proteins and express (or down-regulate) specific genes in plants provides a powerful tool for fundamental research as well as new applications in the field of plant biotechnology. Viable methods that currently exist for protein or gene transfer into plant cells, namely Agrobacterium and microprojectile bombardment, have disadvantages of low transformation frequency, limited host range, or a high cost of equipment and microcarriers. The following protocol outlines a simple and versatile method, which employs rationally-designed peptides as delivery agents for a variety of nucleic acid- and protein-based cargoes into plants. Peptides are selected as tools for development of the system due to their biodegradability, reduced size, diverse and tunable properties as well as the ability to gain intracellular/organellar access. The preparation, characterization and application of optimized formulations for each type of the wide range of delivered cargoes (plasmid DNA, double-stranded DNA or RNA, and protein) are described. Critical steps within the protocol, possible modifications and existing limitations of the method are also discussed.
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spelling pubmed-52264122017-01-26 Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants Chuah, Jo-Ann Horii, Yoko Numata, Keiji J Vis Exp Genetics The capacity to introduce exogenous proteins and express (or down-regulate) specific genes in plants provides a powerful tool for fundamental research as well as new applications in the field of plant biotechnology. Viable methods that currently exist for protein or gene transfer into plant cells, namely Agrobacterium and microprojectile bombardment, have disadvantages of low transformation frequency, limited host range, or a high cost of equipment and microcarriers. The following protocol outlines a simple and versatile method, which employs rationally-designed peptides as delivery agents for a variety of nucleic acid- and protein-based cargoes into plants. Peptides are selected as tools for development of the system due to their biodegradability, reduced size, diverse and tunable properties as well as the ability to gain intracellular/organellar access. The preparation, characterization and application of optimized formulations for each type of the wide range of delivered cargoes (plasmid DNA, double-stranded DNA or RNA, and protein) are described. Critical steps within the protocol, possible modifications and existing limitations of the method are also discussed. MyJove Corporation 2016-12-16 /pmc/articles/PMC5226412/ /pubmed/28060264 http://dx.doi.org/10.3791/54972 Text en Copyright © 2016, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Genetics
Chuah, Jo-Ann
Horii, Yoko
Numata, Keiji
Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title_full Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title_fullStr Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title_full_unstemmed Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title_short Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants
title_sort peptide-derived method to transport genes and proteins across cellular and organellar barriers in plants
topic Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226412/
https://www.ncbi.nlm.nih.gov/pubmed/28060264
http://dx.doi.org/10.3791/54972
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