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

OBJECTIVE: 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 tra...

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Detalles Bibliográficos
Autores principales: Kimura, Mitsuhiro, Endo, Akira, Nagira, Yozo, Yoshizumi, Takeshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10080836/
https://www.ncbi.nlm.nih.gov/pubmed/37024962
http://dx.doi.org/10.1186/s13104-023-06320-3
Descripción
Sumario:OBJECTIVE: 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). RESULTS: 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(2) 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-023-06320-3.