Predation of artificial cells by membrane protein engineering

• Project/Area Number: 15K17454
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Nano/Microsystems
• Research Institution: Kanagawa Academy of Science and Technology
• Principal Investigator: Fujii Satoshi 公益財団法人神奈川科学技術アカデミー, 人工細胞膜システムグループ, 研究員 (60619005)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 人工細胞 / リポソーム / 膜タンパク質 / 膜タンパク質進化工学 / リポソームディスプレイ

Outline of Final Research Achievements

I developed an artificial cells which fuses each together, enabling the “growth” and “predation” of artificial cells. In the present system, the fusogenic activity is encoded in a chimeric gene, by combining a scaffold membrane protein, and a coiled coil domain. This gene was encapsulated in artificial cells and expressed by cell-free translation systems. The expressed proteins were integrated on lipid bilayer, and formed a heterodimer with the protein on the other artificial cells. Consequently, the artificial cells fused together, and the inner content was mixed. As the fusogenic activity is encoded in the gene information, this activity can now be modified by using the in vitro membrane protein evolution techniques.

Research Products

• [Journal Article] Spiral channel for fast and noise-free microRNA detection (2017)
  • Author(s): Fujii Satoshi, Osaki Toshihisa, Misawa Nobuo, Kamiya Koki, Takeuchi Shoji
  • Journal Title: Proc. MEMS 2017 Volume: - Pages: 406-407
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Lipid Bilayer-based Noise-free MicroRNA Detection (2016)
  • Author(s): Fujii Satoshi, Kamiya Koki, Osaki Toshihisa, Misawa Nobuo, Takeuchi Shoji
  • Journal Title: Proc. MicroTAS 2016 Volume: - Pages: 112-113
  • Peer Reviewed
• [Journal Article] Sustainable proliferation of liposomes compatible with inner RNA replication (2016)
  • Author(s): Tsuji, G.; Fujii, S.; Sunami, T.; Yomo, T.
  • Journal Title: Proc. Natl. Acad. Sci. U S A Volume: 113 Issue: 3 Pages: 590-595
  • DOI: 10.1073/pnas.1516893113
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Spiral channel for fast and noise-free microRNA detection (2017)
  • Author(s): Fujii Satoshi, Osaki Toshihisa, Misawa Nobuo, Kamiya Koki, Takeuchi Shoji
  • Organizer: The 30th IEEE International Conference on Micro Electro Mechanical Systems
  • Place of Presentation: アメリカ・ラスベガス
  • Year and Date: 2017-01-22
  • Int'l Joint Research
• [Presentation] Highly efficient formation of droplet interface bilyaers by using a microperforated separator (2017)
  • Author(s): Misawa Nobuo, Fujii Satoshi, Kamiya Koki, Osaki Toshihisa, Takeuchi Shoji
  • Organizer: The 30th IEEE International Conference on Micro Electro Mechanical Systems
  • Place of Presentation: アメリカ・ラスベガス
  • Year and Date: 2017-01-22
  • Int'l Joint Research
• [Presentation] Lipid bilayer at vertically aligned nanoliter droplets generated by two-layered microfluidic channels (2017)
  • Author(s): Toshihisa Osaki, Tomasz Kaminski, Koki Kamiya, Satoshi Fujii, Nobuo Misawa, Piotr Garstecki, Shoji Takeuchi
  • Organizer: The 30th IEEE International Conference on Micro Electro Mechanical Systems
  • Place of Presentation: アメリカ・ラスベガス
  • Year and Date: 2017-01-22
  • Int'l Joint Research
• [Presentation] Electric stress produces bilayer lipid membrane s by exclusion of excessive oil layer (2017)
  • Author(s): Yuya Matsunaga, Toshihisa Osaki, Nobuo Misawa, Satoshi Fujii, Koki Kamiya, Norihisa Miki, Shoji Takeuchi
  • Organizer: The 30th IEEE International Conference on Micro Electro Mechanical Systems
  • Place of Presentation: アメリカ・ラスベガス
  • Year and Date: 2017-01-22
  • Int'l Joint Research
• [Presentation] Lipid Bilayer-based Noise-free MicroRNA Detection (2016)
  • Author(s): Fujii Satoshi, Kamiya Koki, Osaki Toshihisa, Misawa Nobuo, Takeuchi Shoji
  • Organizer: The 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2016)
  • Place of Presentation: アイルランド・ダブリン
  • Year and Date: 2016-10-10
  • Int'l Joint Research
• [Presentation] Odorant Sensor Using An Insect Olfactory Receptor Reconstructed In Artificial Cell Membrane (2016)
  • Author(s): Misawa Nobuo, Fujii Satoshi, Kamiya Koki, Osaki Toshihisa, Miyama Yukiko, Takaku Tomoyuki, Takahashi Yasuhiko, Saito Koichi Takeuchi Shoji
  • Organizer: The 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2016)
  • Place of Presentation: アイルランド・ダブリン
  • Year and Date: 2016-10-10
  • Int'l Joint Research
• [Presentation] Crude Planar Cell Membrane On A Chip (2016)
  • Author(s): Koki Kamiya, Toshihisa Osaki, Kenji.Nakao, Satoshi Fujii, Nobuo Misawa, Shoji Takeuch
  • Organizer: The 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2016)
  • Place of Presentation: アイルランド・ダブリン
  • Year and Date: 2016-10-10
  • Int'l Joint Research
• [Presentation] Multicolored Microcontact Printing Using A Microfluidic Inkpat For Discovery Of Tumor-Imaging Probes In Clinical Specimens (2016)
  • Author(s): Toshihisa Osaki, Haruna Onoyama, Yugo Kuriki, Koki Kamiya, Satoshi Fujii, Nobuo Misawa, Mako Kamiya, Yasuteru Urano, Shoji Takeuchi
  • Organizer: The 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2016)
  • Place of Presentation: アイルランド・ダブリン
  • Year and Date: 2016-10-10
  • Int'l Joint Research
• [Presentation] Biomimetic chemical vapor sensor using agarose gel as mucus (2016)
  • Author(s): Satoshi Fujii, Aiko Nobukawa, Toshihisa Osaki, Nobuo Misawa, Koki Kamiya, Yuya Morimoto, Shoji Takeuchi
  • Organizer: 17th International Symposium on Olfaction and Taste (ISOT2016)
  • Place of Presentation: 日本・神奈川
  • Year and Date: 2016-06-05
  • Int'l Joint Research
• [Presentation] Microrna diagnosis using complementary DNA that brakes transit events through a biological nanopore (2016)
  • Author(s): Toshiya Osaki, Koki Kamiya, Satoshi Fujii, Shoji Takeuchi
  • Organizer: MEMS 2016
  • Place of Presentation: 中国、上海
  • Year and Date: 2016-01-24
  • Int'l Joint Research
• [Presentation] Reconstitution of artificial cell grwoth by coupling the RNA replication and propagation in lipsosomes using a freeze-thaw method (2015)
  • Author(s): Gakushi Tsuji, Satoshi Fujii, Takeshi Sunami, Tetsuya Yomo
  • Organizer: 「細胞を創る」研究会 8.0
  • Place of Presentation: 大阪
  • Year and Date: 2015-11-12
• [Presentation] 人工細胞創造を目指した16S rRNAの進化分子工学的改良 (2015)
  • Author(s): 村瀬由樹、藤井聡志、市橋伯一、角南武志、四方哲也
  • Organizer: 「細胞を創る」研究会 8.0
  • Place of Presentation: 大阪
  • Year and Date: 2015-11-12
• [Presentation] リポソーム膜上に再構成されたEstAのFACSを用いた活性測定 (2015)
  • Author(s): 泉大雅、藤井聡志、市橋伯一、角南武志、四方哲也
  • Organizer: 「細胞を創る」研究会 8.0
  • Place of Presentation: 大阪
  • Year and Date: 2015-11-12
• [Presentation] 単層膜ベシクルへのPEG 脂質の取り込みにおける曲率依存性と閾値 (2015)
  • Author(s): 津田宗一郎、藤井聡志、鈴木宏明、四方哲也
  • Organizer: 「細胞を創る」研究会 8.0
  • Place of Presentation: 大阪
  • Year and Date: 2015-11-12
• [Presentation] 膜タンパク質を人工細胞に組み込み、進化させる (2015)
  • Author(s): Satoshi Fujii
  • Organizer: 第５３回日本生物物理学会年回
  • Place of Presentation: 金沢
  • Year and Date: 2015-09-13
  • Invited
• [Presentation] Freeze thaw method for supply small nutrients to the liposome by inducing liposome fusion (2015)
  • Author(s): Gakushi Tsuji, Satoshi Fujii, Takeshi Sunami, Tetsuya Yomo
  • Organizer: 第53回日本生物物理学会
  • Place of Presentation: 金沢
  • Year and Date: 2015-09-13
• [Book] 膜タンパク質とDNAを用いたバイオセンサ (2017)
  • Author(s): 藤井聡志、竹内昌治
  • Total Pages: 8
  • Publisher: 化学工業社
• [Book] 人工細胞膜を利用した1分子検出 (2016)
  • Author(s): 藤井聡志、神谷厚輝、竹内昌治
  • Total Pages: 5
  • Publisher: 東京化学同人
• [Patent(Industrial Property Rights)] 核酸の検出方法及び検出装置 (2016)
  • Inventor(s): 藤井聡志 神谷厚輝 大崎寿久 三澤宣雄 竹内昌治
  • Industrial Property Rights Holder: （公財）神奈川科学技術アカデミー
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-10-07