Plant Biotechnology
Online ISSN : 1347-6114
Print ISSN : 1342-4580
ISSN-L : 1342-4580
A centrifugation-assisted peptide-mediated gene transfer method for high-throughput analyses
Mitsuhiro KimuraTakeshi YoshizumiKeiji Numata
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JOURNALS FREE ACCESS Advance online publication

Article ID: 18.1115a

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Abstract

A peptide-mediated DNA delivery system for several plant species has been recently developed. This system uses ionic complexes of DNA and fusion peptides containing several domains, such as DNA-binding and cell-penetrating peptides. Although the peptide-DNA complexes are capable of penetrating into plant cells through the cell wall by mechanical pressure using a syringe, sample throughput is limited. Here, we describe a Centrifugation-Assisted Peptide-mediated gene Transfer (CAPT) method for improving sample throughput with reproducible gene transfer efficiency. We optimized the parameters of CAPT for transient gene transfer efficiency by using Nicotiana tabacum cotyledons as a model plant material. The optimal parameters for centrifugation were 10,000×g for 60 s. Furthermore, we successfully transferred the peptide–DNA complex into rice cotyledons using the optimized CAPT method. Thus, the CAPT method is superior to the previous syringe-mediated infiltration method in terms of sample throughput in multiple plant species.

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© 2019 The Japanese Society for Plant Cell and Molecular Biology
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