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 publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6435
DOIs
Publication statusPublished - 2007
EventOptical Interactions with Tissue and Cells XVIII - San Jose, CA, United States
Duration: 2007 Jan 222007 Jan 24

Other

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

Fingerprint

Liposomes
DNA
Genes
Gene transfer
Lasers
Laser pulses
Fluorescence
Catheters
Cell membranes
Charge density
Controllability
Cell culture
Rubber
Microscopes
Irradiation
Proteins

ASJC Scopus subject areas

  • Engineering(all)

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 Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 6435). [643518] https://doi.org/10.1117/12.701734

Laser induced stress wave-assisted gene transfection : Improved transfection efficiency with cationic liposome-modified plasmid DNA. / Otsuka, Risa; Terakawa, Mitsuhiro; Sato, Shunichi; Satoh, Yasushi; Takishima, Kunio; Ashida, Hiroshi; Obara, Minoru.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6435 2007. 643518.

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

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 Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 6435, 643518, Optical Interactions with Tissue and Cells XVIII, San Jose, CA, United States, 07/1/22. https://doi.org/10.1117/12.701734
Otsuka R, Terakawa M, Sato S, Satoh Y, Takishima K, Ashida H et al. Laser induced stress wave-assisted gene transfection: Improved transfection efficiency with cationic liposome-modified plasmid DNA. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6435. 2007. 643518 https://doi.org/10.1117/12.701734
Otsuka, Risa ; Terakawa, Mitsuhiro ; Sato, Shunichi ; Satoh, Yasushi ; Takishima, Kunio ; Ashida, Hiroshi ; Obara, Minoru. / Laser induced stress wave-assisted gene transfection : Improved transfection efficiency with cationic liposome-modified plasmid DNA. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6435 2007.
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