Particle bombardment-assisted peptide-mediated gene transfer for highly efficient transient assay

• Published in: BMC research notes Vol. 16; no. 1; p. 46
• Main Authors: Kimura, Mitsuhiro, Endo, Akira, Nagira, Yozo, Yoshizumi, Takeshi
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 06.04.2023; BioMed Central; BMC
• Subjects: Centrifugation; Chloroplasts; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA - genetics; DNA - metabolism; DNA-peptide complex; Efficiency; Flowers & plants; Gene expression; Gene transfer; Gene Transfer Techniques; Genes; Genomes; Gold; Membranes; Mitochondria; Mitochondrial DNA; Nanoluc; Particle bombardment; Particle bombardment-assisted peptide-mediated gene transfer (PBPT); Peptides; Peptides - genetics; Permeability; Plant cells; Plant genetics; Plant mitochondria; Plants; Research Note; Seeds; Transformation, Genetic; Japan; United States > US; Nanoluc; Particle bombardment-assisted peptide-mediated gene transfer (PBPT); DNA-peptide complex
• ISSN: 1756-0500; 1756-0500
• DOI: 10.1186/s13104-023-06320-3

A centrifugation-assisted peptide-mediated gene transfer (CAPT) method was recently developed as an efficient system for gene delivery into plant cells. However, the gene transfer efficiency of CAPT into plant cells was not entirely satisfactory for detecting transient expression of a transgene driven into mitochondria. Here, we report a new gene delivery system using a method called particle bombardment-assisted peptide-mediated gene transfer (PBPT). We investigated various parameters of the PBPT method to increase transient gene expression efficiency in Brassica campestris. The optimal conditions for PBPT were a single bombardment with gold particles coated with a DNA‒peptide complex (6 µg of DNA and 2 µg of peptide) at an acceleration pressure of 5 kg/cm and a target distance of 12.5 cm. Moreover, bombardment under the optimal conditions successfully transferred the transgene into the cells of other plant species, namely B. juncea and tomato. Thus, we developed a PBPT method for highly efficient delivery of a DNA‒peptide complex into plant mitochondria.