Gene transfer experiments by using a combination of electric pulses

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K06332
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Power engineering/Power conversion/Electric machinery
• Research Institution: Ariake National College of Technology
• Principal Investigator: Kono Susumu 有明工業高等専門学校, 創造工学科, 教授 (30270375)
• Co-Investigator(Kenkyū-buntansha): 冨永 伸明 有明工業高等専門学校, 創造工学科, 教授 (30227631)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: パルスパワー / ナノ秒 / 遺伝子導入 / GFP / HL60

Outline of Final Research Achievements

In electroporation, voltage pulses are used to create pores on cell membrane, and then genes are expected to be introduced into cells through the pores. A condition of voltage pulse is very important factor to achieve high efficiency of introduction.
In this study, rectangular voltage pulses with wide range from nano-second to a second were applied to cells for increasing the membrane permeability.
Two hundred nanosecond pulse seldom used in ordinary electroporators was found to increase the membrane permeability with lower energy input than ordinary voltage pulses. Combination of the 200 ns pulse as 1st pulse and the low voltage pulse (4 V - 400 V and 10 microsecond - 1 second) as 2nd pulse were tested. The outcomes of transformation efficiencies were posted in a graph with an electric field on the perpendicular axis and an input energy on the horizontal axis. The pulse parameters to achieve high efficiency of introduction were easily found by using this graph.

Free Research Field

パルスパワー

Academic Significance and Societal Importance of the Research Achievements

従来のエレクトロポレーション法による遺伝子導入では導入が困難である細胞が依然として存在しており，その導入効率の向上が望まれている。
今回の研究結果から，ナノ秒高電圧パルスに低電圧長パルスを組み合わせることで，導入困難と考えられている数種類の細胞への遺伝子導入が可能であることが分かった。これらのパルスを組み合わせた遺伝子導入実験により，効率の良い遺伝子導入が達成できるパルス条件を簡便に見出すことができる手法を提案した。
本手法を適用することで，現在遺伝子導入が困難とされる細胞に対する導入効率が向上すれば，医学，薬学，生物学などの幅広い分野に貢献できる技術になると考えられる。