Fast multiplex calcium recording using a red-shift calcium indicator gene

• Project/Area Number: 16K13105
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Basic / Social brain science
• Research Institution: The University of Tokyo
• Principal Investigator: Bito Haruhiko 東京大学, 大学院医学系研究科(医学部), 教授 (00291964)
• Project Period (FY): 2016-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2016: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
• Keywords: カルシウム / シグナル伝達 / 脳・神経

Outline of Final Research Achievements

We previously developed a genetically encoded calcium indicator RCaMP2, which surpasses performance obtained with classical genetically encoded or chemical dye-based calcium indicators. Transformative insights for rational design of R-CaMP2 were gained from previous biochemical research on calcium-calmodulin-binding proteins pioneered by the investigator group. In this study, we expanded this strategy, and rationally pursued protein engineering-based iterative mutation processes to realize multiple color calcium recording that goes beyond the classical two colors, green and red. Multicolor cellular activiy measurements represent an urgent technological problem to solve. Our approach opens a hitherto unexplored, brand-new, avenue for a novel fast multiplex calcium imaging platform through an original GECI design concept.

Research Products

• [Journal Article] Calmodulin kinases: essential regulators in health and disease. (2017)
  • Author(s): Takemoto-Kimura S, Suzuki K, Horigane SI, Kamijo S, Inoue M, Sakamoto M, Fujii H, Bito H.
  • Journal Title: J Neurochem. Volume: - Issue: 6 Pages: 808-818
  • DOI: 10.1111/jnc.14020
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Simultaneous fast measurement of circuit dynamics at multiple sites across the mammalian brain. (2016)
  • Author(s): Kim CK, Yang SJ, Pichamoorthy N, Young NP, Kauvar I, Jennings JH, Lerner TN, Berndt A, Lee SY, Ramakrishnan C, Davidson TJ, Inoue M, Bito H, Deisseroth K.
  • Journal Title: Nature Methods Volume: 13 Issue: 4 Pages: 325-328
  • DOI: 10.1038/nmeth.3770
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Multiplex imaging of neural activity and signaling dynamics. (2016)
  • Author(s): Bito H.
  • Organizer: Janelia Conferences - Fall 2016 “Fluorescent Proteins and Biological Sciences V”.
  • Place of Presentation: Janelia Farm Research Campus, Ashburn, Virginia, USA
  • Year and Date: 2016-11-06
  • Int'l Joint Research / Invited