Micro and nano electroporation for direct delivery of molecules to cell nucleus

2018 Fiscal Year Final Research Report

• Project/Area Number: 26289035
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Fluid engineering
• Research Institution: Institute of Physical and Chemical Research (2018); Kyoto University (2014-2017)
• Principal Investigator: Shintaku Hirofumi 国立研究開発法人理化学研究所, 開拓研究本部, 理研白眉研究チームリーダー (80448050)
• Co-Investigator(Kenkyū-buntansha): 小寺 秀俊 京都大学, 工学研究科, 教授 (20252471)
• Project Period (FY): 2014-04-01 – 2019-03-31
• Keywords: RNA / DNA / 電気泳動 / マイクロ流体システム / シーケンシング / 1細胞 / 電気穿孔

Outline of Final Research Achievements

We developed a microfluidic system that offered a highly reproducible single-cell isolation and electroporation (97%, n>120). We showed that the hydrodynamic trap effectively captured a single cell and focused the electric field for electroporation to reduce the applied voltage by ten-fold in comparison to a uniform electric field. We also developed an equation that estimated the magnitude of the electric field exerted on the nuclear membrane as a function of the distance between the hydrodynamic trap and the nuclear membrane. Furthermore, we applied our microfluidic system for integrated nuclear and cytoplasmic RNA-sequencing of single cells and coined the method SINC-seq.

Free Research Field

マイクロ流体工学

Academic Significance and Societal Importance of the Research Achievements

本研究で開発したマイクロ流体システムは集中電場を活用して選択的に細胞膜を穿孔し，細胞内外の物質輸送を制御することを実現した．これを活用して，再現性の高い電気穿孔法を確立した．さらに，世界で初めて1細胞の細胞質RNAと核RNAを独立かつ並列でシーケンス解析するSINC-seq法(1細胞RNA分画解読法)を確立した．