Laser induced stress wave-assisted gene transfection: Improved transfection efficiency with cationic liposome-modified plasmid DNA

Risa Otsuka, Mitsuhiro Terakawa, Shunichi Sato, Yasushi Satoh, Kunio Takishima, Hiroshi Ashida, Minoru Obara

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

    Abstract

    Although various methods for gene transfer have been investigated, a practical gene delivery system that fulfills the requirements for clinical application has not yet been developed. Gene transfer by the use of laser-induced stress waves (LISWs) is a physical method to facilitate gene transfer into cells with the effect of stress waves generated by irradiation an absorbing material with high-intensity laser pulses. This method has high spatial controllability and a potential for catheter-based gene transfer. We demonstrated selective high transfection efficiency in vivo. However, there remains a problem that transfection efficiency is limited to less than several percent in vitro. Thus we attempted to improve transfection efficiency by using plasmid DNA modified with cationic-liposome. Plasmid DNA coding for enhanced green fluorescent protein (EGFP) had been modified with Lipofectamine and it was added to a dish for NIH3T3 cell culture. A black rubber disk was placed on the upper side of the cells; the disk was irradiated with 532 nm, nanosecond laser pulses (spot diameter, 3 mm; fluence, 1.3 J/cm2; number of pulses, 20). 24 hours after application of LISWs, transfection efficiency was evaluated with a fluorescence microscope, where efficiency was defined as the ratio of the number of cells emitting fluorescence to the total number of cells. At a DNA concentration of 7.8 μg/ml, transfection efficiency with naked plasmid DNA was as low as 0.05%, while it was increased to 23.7% by using plasmid DNA modified with Lipofectamine. Since both of the naked plasmid DNA and cell membranes have negative charge, plasmid DNA concentration around cells should be low. Since DNA-Lipofectamine complexes carry positive charge, density of plasmids existing around cells should be increased, resulting in much improved transfection efficiency.

    Original languageEnglish
    Title of host publicationOptical Interactions with Tissue and Cells XVIII
    DOIs
    Publication statusPublished - 2007 Apr 30
    EventOptical Interactions with Tissue and Cells XVIII - San Jose, CA, United States
    Duration: 2007 Jan 222007 Jan 24

    Publication series

    NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
    Volume6435
    ISSN (Print)1605-7422

    Other

    OtherOptical Interactions with Tissue and Cells XVIII
    CountryUnited States
    CitySan Jose, CA
    Period07/1/2207/1/24

    ASJC Scopus subject areas

    • Electronic, Optical and Magnetic Materials
    • Biomaterials
    • Atomic and Molecular Physics, and Optics
    • Radiology Nuclear Medicine and imaging

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  • Cite this

    Otsuka, R., Terakawa, M., Sato, S., Satoh, Y., Takishima, K., Ashida, H., & Obara, M. (2007). Laser induced stress wave-assisted gene transfection: Improved transfection efficiency with cationic liposome-modified plasmid DNA. In Optical Interactions with Tissue and Cells XVIII [643518] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 6435). https://doi.org/10.1117/12.701734