High-throughput analysis of olfactory receptor repertoire by using the automated single-cell analysis and isolation system

• Project/Area Number: 15K21068
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied molecular and cellular biology; Cell biology
• Research Institution: Osaka University
• Principal Investigator: Yoshimoto Nobuo 大阪大学, 産業科学研究所, 特任准教授 (80467612)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: １細胞解析 / 嗅覚細胞 / 嗅覚受容体 / 匂い分子 / 嗅覚受容体レパトア / バイオセンサー / 全自動１細胞解析単離装置 / タイムラプスサイトメトリー

Outline of Final Research Achievements

A cell array consisting of 200,000 olfactory epithelial cells was formed on a microchamber array chip enabling cell alignment. After installing this cell array chip in the automated single-cell analysis and isolation system equipped with a perfusion system, the cells were subjected to time-lapse cytometry. Olfactory cells which transiently responded to stimulation of specific odor molecules were identified, and then the desired olfactory cells were isolated at single cell level. Each isolated single cell was subjected to genetic analysis to identify a combination of olfactory receptor groups that bind odor molecules specifically.

Research Products

• [Journal Article] High-Throughput Analysis of Mammalian Receptor Tyrosine Kinase Activation in Yeast Cells (2016)
  • Author(s): Nobuo Yoshimoto, Shun'ichi Kuroda
  • Journal Title: Methods in Molecular Biology pp Volume: 1487 Pages: 35-52
  • DOI: 10.1007/978-1-4939-6424-6_3
  • ISBN: 9781493964222, 9781493964246
  • Peer Reviewed / Open Access
• [Journal Article] Cellular uptake of hepatitis B virus envelope L particles is independent of sodium taurocholate cotransporting polypeptide, but dependent on heparan sulfate proteoglycan (2016)
  • Author(s): Masaharu Somiya, Qiushi Liu, Nobuo Yoshimoto, Masumi Iijima, Kenji Tatematsu, Tadashi Nakai, Toshihide Okajima, Kazuyuki Kuroki, Keiji Ueda, Shun’ichi Kuroda
  • Journal Title: Virology Volume: 497 Pages: 23-32
  • Peer Reviewed / Open Access
• [Journal Article] RBM20 and RBM24 cooperatively promote the expression of short enh splice variants (2016)
  • Author(s): Jumpei Ito, Masumi Iijima, Nobuo Yoshimoto, Tomoaki Niimi, Shun'ichi Kuroda, Andres D. Maturana
  • Journal Title: FEBS Letter Volume: 590 Issue: 14 Pages: 2262-2274
  • DOI: 10.1002/1873-3468.12251
  • Peer Reviewed / Open Access
• [Journal Article] Mutational analysis of hepatitis B virus pre-S1 (9-24) fusogenic peptide (2016)
  • Author(s): Qiushi Liu, Masaharu Somiya, Naohiko Shimada, Wakako Sakamoto, Nobuo Yoshimoto, Masumi Iijima, Kenji Tatematsu, Tadashi Nakai, Toshihide Okajima, Atsushi Maruyama, Shun'ichi Kuroda
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 474 Issue: 2 Pages: 406-412
  • DOI: 10.1016/j.bbrc.2016.04.125
  • Peer Reviewed / Open Access
• [Journal Article] Scaffold protein enigma homolog 1 overcomes the repression of myogenesis activation by inhibitor of DNA binding 2 (2016)
  • Author(s): Nakatani M. Ito J. Koyama R. Iijima M. Yoshimoto N. Niimi T. Kuroda S. Maturana AD
  • Journal Title: Biochem Biophys Res Commun. Volume: 0 Issue: 2 Pages: 0-0
  • DOI: 10.1016/j.bbrc.2016.04.119
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Bio-nanocapsule-based scaffold improves the sensitivity and ligand-binding capacity of mammalian receptors on the sensor chip (2016)
  • Author(s): Iijima, M., Yoshimoto, N., Niimi, T., Maturana, A.D., Kuroda, S.
  • Journal Title: Biotechnology journal Volume: 印刷中 Issue: 6 Pages: 805-813
  • DOI: 10.1002/biot.201500443
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Bio-nanocapsules displaying various immunoglobulins as an active targeting-based drug delivery system (2016)
  • Author(s): Kenji Tatematsu, Masumi Iijima, Nobuo Yoshimoto, Tadashi Nakai, Toshihide Okajima, Shun’ichi Kuroda
  • Journal Title: Acta Biomaterialia Volume: 35 Pages: 238-247
  • DOI: 10.1016/j.actbio.2016.02.010
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Cytokine-dependent activation of the JAK-STAT pathway in Saccharomyces cerevisiae (2016)
  • Author(s): Yoshimoto, N., Ikeda, Y., Tatematsu, K., Iijima, M., Nakai, T., Okajima, T., Tanizawa, K., Kuroda, S.
  • Journal Title: Biotechnology and Bioengineering Volume: 印刷中 Issue: 8 Pages: 1796-1804
  • DOI: 10.1002/bit.25948
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Deciphering the Receptor Repertoire Encoding Specific Odorants by Time-Lapse Single-Cell Array Cytometry (2016)
  • Author(s): Suzuki M, Yoshimoto N, Shimono K, Kuroda S
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 19934-19934
  • DOI: 10.1038/srep19934
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] High-throughput analysis of mammalian receptor tyrosine kinase activation in yeast cells (2016)
  • Author(s): Yoshimoto N, Kuroda S
  • Journal Title: Methods in Molecular Biology Volume: 9999
  • Peer Reviewed
• [Journal Article] Mapping the heparin-binding site of the osteoinductive protein NELL1 by site-directed mutagenesis. (2015)
  • Author(s): Takahashi, K., Imai, A., Iijima, M., Yoshimoto, N., Maturana, A. D., Kuroda, S., and Niimi, T.
  • Journal Title: FEBS Letters Volume: 589 Issue: 24PartB Pages: 4026-4032
  • DOI: 10.1016/j.febslet.2015.11.032
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Scaffold protein enigma homolog activates CREB whereas a short splice variant prevents CREB activation in cardiomyocytes (2015)
  • Author(s): Ito J, Iijima M, Yoshimoto N, Niimi T, Kuroda S, Maturana AD
  • Journal Title: Cellular Signaling Volume: 27 Issue: 12 Pages: 2425-2433
  • DOI: 10.1016/j.cellsig.2015.09.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy (2015)
  • Author(s): Liu Q, Jung J, Somiya M, Iijima M, Yoshimoto N, Niimi T, Maturana AD, Shin SH, Jeong SY, Choi EK, Kuroda S
  • Journal Title: International Journal of Nanomedicine Volume: 10 Pages: 4159-4172
  • DOI: 10.2147/ijn.s84295
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Intracellular trafficking of bio-nanocapsule-liposome complex: Identification of fusogenic activity in the pre-S1 region of hepatitis B virus surface antigen L protein (2015)
  • Author(s): Somiya M, Sasaki Y, Matsuzaki T, Liu Q, Iijima M, Yoshimoto N, Niimi T, Maturana AD, Kuroda S
  • Journal Title: Journal of Controlled Release Volume: 212 Pages: 10-18
  • DOI: 10.1016/j.jconrel.2015.06.012
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] One-step scalable preparation method for non-cationic liposomes with high siRNA content (2015)
  • Author(s): Somiya M, Yamaguchi K, Liu Q, Niimi T, Maturana AD, Iijima M, Yoshimoto N, Kuroda S
  • Journal Title: International Journal of Pharmaceutics Volume: 490 Issue: 1-2 Pages: 316-323
  • DOI: 10.1016/j.ijpharm.2015.05.072
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Decipherment of olfactory receptor repertoire by using an automated single-cell analysis and isolation system (2016)
  • Author(s): Nobuo Yoshimoto, Masato Suzuki, Akiko Kida, Ken Shimono, Shun'ichi Kuroda
  • Organizer: The 20th ISIR International Symposium
  • Place of Presentation: 大阪府大阪市
  • Year and Date: 2016-12-12
  • Int'l Joint Research
• [Presentation] 細胞表層FIAとイムノチャンバーを用いる１細胞育種法の開発 (2016)
  • Author(s): 良元伸男、黒田俊一
  • Organizer: 第68回日本生物工学会大会
  • Place of Presentation: 富山県富山市
  • Year and Date: 2016-09-28
• [Presentation] Decipherment of olfactory receptor repertoire by using an automated single-cell analysis and isolation system equipped with real-time calcium imaging device (2015)
  • Author(s): Nobuo Yoshimoto
  • Organizer: Pacifichem 2015
  • Place of Presentation: Honolulu (USA)
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] 全自動１細胞解析単離装置を用いる１細胞育種とその応用技術 (2015)
  • Author(s): 良元伸男、木田晶子、黒田俊一
  • Organizer: 第67回日本生物工学会大会
  • Place of Presentation: 城山観光ホテル（鹿児島県鹿児島市）
  • Year and Date: 2015-10-26
• [Presentation] 全自動１細胞解析単離装置による１細胞育種と様々な応用成果 (2015)
  • Author(s): 良元伸男
  • Organizer: 2015年度三賞受賞者合同発表会 in BioJapan2015
  • Place of Presentation: パシフィコ横浜アネックスホール（神奈川県横浜市）
  • Year and Date: 2015-10-14
  • Invited
• [Presentation] 全自動１細胞解析単離装置を用いる優良細胞の１細胞育種 (2015)
  • Author(s): 良元伸男、木田晶子、黒田俊一
  • Organizer: 第28回 日本動物細胞工学会2015年度大会 (JAACT2015)
  • Place of Presentation: 東北大学 片平さくらホール（宮城県仙台市）
  • Year and Date: 2015-07-09
• [Presentation] ヒト化抗体高分泌CHO細胞の１細胞スクリーニング法の開発 (2015)
  • Author(s): 木田晶子、良元伸男、黒田俊一
  • Organizer: 第28回 日本動物細胞工学会2015年度大会 (JAACT2015)
  • Place of Presentation: 東北大学 片平さくらホール（宮城県仙台市）
  • Year and Date: 2015-07-09
• [Book] AROMA RESEARCH No.67 「特定の匂い分子に応答する嗅覚受容体群の網羅的取得法」 (2016)
  • Author(s): 良元伸男、黒田俊一
  • Total Pages: 6
  • Publisher: フレグランスジャーナル
• [Book] バイオサイエンスとインダストリー「【解説】１細胞育種を実現する全自動1 細胞解析単離装置の開発」 (2016)
  • Author(s): 良元伸男、黒田俊一
  • Total Pages: 4
  • Publisher: バイオインダストリー協会
• [Book] ファインケミカルシリーズ 抗体医薬における細胞構築・培養・ダウンストリームのすべて「第５章 １細胞育種を実現する全自動１細胞解析単離措置の開発」 (2015)
  • Author(s): 良元伸男、黒田俊一
  • Total Pages: 11
  • Publisher: シーエムシー出版
• [Remarks] 生体分子反応科学研究室
  • URL: http://www.sanken.osaka-u.ac.jp/labs/smb/index.html