New Technique for Gene Transfection Using Laser Irradiation

Shirahata, Yasuhiro; Ohkohchi, Nobuhiro; Itagak, Hideo; Satoh, Susumu

doi:10.2310/6650.2001.34045

Cited by 65 publications

(30 citation statements)

References 19 publications

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“…It has been previously shown that the transfection efficiency decreases significantly when transfected with a plasmid concentration less than 10 mg/ml [3,15]. However we show that, in the presence of Nupherin-neuron, we could lower the plasmid concentration to 3.3 mg/ml without compromising the resulting transfection efficiency.…”

Section: Biophotonicscontrasting

confidence: 55%

Enhancement and optimization of plasmid expression in femtosecond optical transfection

Praveen

Stevenson

Antkowiak

et al. 2011

Journal of Biophotonics

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Cell transfection using femtosecond lasers is gaining importance for its proven ability to achieve selective transfection in a sterile and relatively non‐invasive manner. However, the net efficiency of this technique is limited due to a number of factors that ultimately makes it difficult to be used as a viable and widely used technique. We report here a method to achieve significant enhancement in the efficiency of femtosecond optical transfection. The transfection procedure is modified by incorporating a suitable synthetic peptide containing nuclear localization and DNA binding sequences, assisting DNA import into the nucleus. We achieved a 3‐fold enhancement in the transfection efficiency for adherent Chinese Hamster Ovary (CHO‐K1) cells with this modified protocol. Further, in the presence of this biochemical reagent, we were able to reduce the required plasmid concentration by ∼70% without compromising the transfection efficiency. Also, we report for the first time the successful photo‐transfection of recently trypsinised cells with significantly high transfection efficiency when transfected with modified plasmid. This paves the way for the development of high throughput microfluidic optical transfection devices. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Section: Biophotonicscontrasting

confidence: 55%

Enhancement and optimization of plasmid expression in femtosecond optical transfection

Praveen

Stevenson

Antkowiak

et al. 2011

Journal of Biophotonics

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“…Other studies of laser-induced gene delivery were carried out on single cells. [8][9][10][11] Our results, that laser engery is capable of enhancing gene transfer as well in cell suspensions, are promising for future in vivo laser application in the prostate. James et al 14 considered that direct in vivo gene transfer to urological organs is possible by physical methods like electroporation.…”

Section: Discussionmentioning

confidence: 64%

“…They also noted that the permeability of the cell membrane is modified at the site of the beam impact. Shirahata et al 10 observed a similar effect of gene transfer when a pulse laser irradiation was focused on a cell membrane. All the above studies show that the cells upon laser irradiation undergo a change in the permeability of the plasma membrane or form pores in the membrane at the site of contact.…”

Section: Introductionmentioning

confidence: 84%

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Gene delivery into prostate cancer cells by holmium laser application

Sagi

Knoll

Trojan

et al. 2003

Prostate Cancer Prostatic Dis

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New approaches to treat prostate cancer (PCA) are utilizing gene therapy and aim to correct the disease at the genetic level. Getting a gene efficiently into the target cell is the subject of much interest. We used a holmium laser for transfecting rat PCA cells with the reporter gene pEGFP. By FACS analysis and fluorescence microscopy, we could demonstrate that cellular delivery of plasmid DNA was possible with high efficiencies up to 41.3%. Therefore, transfection of PCA cells by holmium laser might offer a promising new gene transfer strategy to PCA with minimal invasiveness.

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“…Optical cell translocation satisfies these criteria. A variety of lasers, ranging from continuous wave visible sources [4][5][6][7][8] to pulsed infrared [9][10][11][12] sources, have been applied for efficient introduction of biomaterials into mammalian cells.…”

Section: Introductionmentioning

confidence: 99%