2021 Fiscal Year Annual Research Report
Directed Evolution of a Palette of Optogenetic and Chemi-Optogenetic Indicators for Multiplexed Imaging of Cellular Metabolism
| Project/Area Number |
19H05633
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| Research Institution | The University of Tokyo |
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Principal Investigator |
キャンベル ロバート.アール 東京大学, 大学院理学系研究科(理学部), 教授 (40831318)
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| Co-Investigator(Kenkyū-buntansha) |
合田 圭介 東京大学, 大学院理学系研究科(理学部), 教授 (70518696)
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| Project Period (FY) |
2019-06-26 – 2024-03-31
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| Keywords | Protein Engineering / Fluorescence / Microscopy / Cell Biology |
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| Outline of Annual Research Achievements |
In FY2020 we established a number of new designs for new genetically encoded biosensors. In FY2021 our efforts focussed on their optimization, characterization, and collaborative applications. Our flagship project is a biosensor of lactate. In FY2021, we published the design, optimization, characterization and application of the first generation green fluorescent biosensor in Nature Communications. Many other projects in the lab are following a similar trajectory, although they are at earlier stages. Among these projects are efforts to develop red and green biosensors of intracellular and extracellular lactate, and near-infrared Ca2+ biosensors. We expect these projects to be published in FY2022. Other projects at earlier development stages include new biosensors for metabolites such as pyruvate, fumarate, and citrate. Our projects in the area of chemigenetic biosensors are following a parallel trajectory to the projects on fully genetically encoded biosensors. In FY2020, we had developed a new design of HaloTag based biosensor that could be applied for either Ca2+ or Na+, and a PYP-based biosensor for Ca2+. During FY2021 we have been focussed on their further development. We expect that these biosensor to be published in FY2022 or FY2023. Other projects at earlier development stages include the development of near-infrared fluorescent chemigenetic biosensors for Ca2+ and K+. In FY2021, Campbell gave 8 invited seminars about this research, including ones at University of British Columbia, the China-Japan-Korea Symposium, and Photonics North (keynote; online).
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
In the area of fully genetically encoded biosensors, we were able to report our first generation biosensors of extracellular lactate (Nat. Commun., 2021, 12, 7058). We also made great progress in the development of new NIR calcium sensors. In the area of chemi-optogenetic biosensors, we made tremendous progress in FY2021 and our first manuscript is current under revision for Nature Chemical Biology, having received consistently strong reviews.
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| Strategy for Future Research Activity |
Our research plan for FY2022 and beyond is to continue the development of various genetically encoded and chemi-optogenetic biosensors for a range of metabolites, inorganic ions, and various molecules of biological interest.
In the area of fully genetically encoded biosensors, we have recently reported our first generation biosensors of extracellular lactate. Work on the second generation extracellular lactate biosensors is nearing completion and likely to be published during FY2022. In parallel, we will continue work on our first generation green and red fluorescent intracellular lactate biosensors, which is likely to be published in FY2022. We are particularly excited about exploring multicolour, multiplexed imaging of intra- and extra-cellular lactate concentrations using strategic combinations of green and red fluore
scent biosensors.
In the area of chemi-optogenetic biosensors, during FY2022, we will build upon the success of the project in FY2021 and develop a variety of other chemi-optogenetic sensors, including some that are HaloTag-based and some that are
PYP-based.
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[Journal Article] Deep imaging flow cytometry2022
Author(s)
Huang Kangrui、Matsumura Hiroki、Zhao Yaqi、Herbig Maik、Yuan Dan、Mineharu Yohei、Harmon Jeffrey、Findinier Justin、Yamagishi Mai、Ohnuki Shinsuke、Nitta Nao、Grossman Arthur R.、Ohya Yoshikazu、Mikami Hideharu、Isozaki Akihiro、Goda Keisuke
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Journal Title
Lab on a Chip
Volume: 22
Pages: 876～889
DOI
Peer Reviewed / Int'l Joint Research
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[Journal Article] Analyzing circulating platelet aggregates in patients with COVID-19 by massive image-based single-cell profiling2022
Author(s)
W. Peterson, J. Takiguchi, M. Shifat-E-Rabbi, Y. Zhuang, X. Yin, A. H. M. Rubaiyat, Y. Deng, H. Zhang, S. Miyata, G. K. Rohde, W. Iwasaki, Y. Yatomi, and K. Goda
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Journal Title
化学とマイクロ・ナノシステム学会誌
Volume: 21
Pages: 1～11
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[Presentation] High-throughput on-chip imaging of diverse cancer cells in blood2021
Author(s)
Hiroki Matsumura, Larina Tzu-Wei Shen, Hideharu Mikami, Akihiro Isozaki, Dan Yuan, Taichi Miura, Yoshika Kusumoto, Masako Nishikawa, Atsushi Yasumoto, Takeshi Yamada, Aya Ueda, Hiroko Bando, Hisato Hara, Yutaka Yatomi, Satoshi Matsusaka, and Keisuke Goda
Organizer
Pacifichem 2021
Int'l Joint Research
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