Visualization of cell fates using degron based fluorescence probes.

• Project/Area Number: 25440095
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: SAKAUE ASAKO 国立研究開発法人理化学研究所, 脳科学総合研究センター, 研究員 (90462689)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 多細胞社会 / 生命現象可視化 / デグロン / 蛍光タンパク / Fucci / Ｆｕｃｃｉ / 蛍光性プローブ / 可視化 / 運命決定スイッチ / 蛍光プローブ

Outline of Final Research Achievements

To understand the spatio-temporal dynamics of cellular functions in complex multi-cellular systems, we have developed a variety of fluorescent indicators for live cell imaging. The indicators include a Fucci probe that resolves a cell cycle into 7 fractions (CC (Cell-Cycle) Rainbow), oxidative stress probes, and carbohydrate metabolism probes. Practical use of these probes gave us opportunities to realize “cell heterogeneity” and appreciate “cell personality” in many cell culture systems. Our forthcoming studies will focus on generation of flox mouse lines of the probes that allow us to better understand the spatio-temporal regulations of cellular functions in a more physiological context.

Research Products

• [Journal Article] Involvement of Receptor Activator of Nuclear Factor-κB Ligand (RANKL)-induced Incomplete Cytokinesis in the Polyploidization of Osteoclasts. (2016)
  • Author(s): Takegahara N, Kim H, Mizuno H, Sakaue-Sawano A, Miyawaki A, Tomura M, Kanagawa O, Ishii M, Choi Y.
  • Journal Title: The Journal of Biological Chemistry Volume: 291 Issue: 7 Pages: 3439-3454
  • DOI: 10.1074/jbc.m115.677427
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Proliferation-coupled osteoclast differentiation by RANKL: Cell density as a determinant of osteoclast formation. (2015)
  • Author(s): Motiur Rahman M, Takeshita S, Matsuoka K, Kaneko K, Naoe Y, Sakaue-Sawano A, Miyawaki A, Ikeda K.
  • Journal Title: Bone. Volume: 81 Pages: 392-399
  • DOI: 10.1016/j.bone.2015.08.008
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Fucci-guided purification of hematopoietic stem cells with high repopulating activity. (2015)
  • Author(s): Yo M, Sakaue-Sawano A, Noda S, Miyawaki A, *Miyoshi H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 457 Issue: 1 Pages: 7-11
  • DOI: 10.1016/j.bbrc.2014.12.074
  • Peer Reviewed
• [Journal Article] Real-time tracking of cell cycle progression during CD8(+) effector and memory T-cell differentiation. (2015)
  • Author(s): Kinjyo I, Qin J, Tan SY, Wellard CJ, Mrass P, Ritchie W, Doi A, Cavanagh LL, Tomura M, Sakaue-Sawano A, Kanagawa O, Miyawaki A, Hodgkin PD, Weninger W.
  • Journal Title: Nat Commun. Volume: 6 Issue: 1 Pages: 6301-6301
  • DOI: 10.1038/ncomms7301
  • Peer Reviewed / Open Access
• [Journal Article] Visualizing the cell-cycle progression of endothelial cells in zebrafish. (2014)
  • Author(s): Fukuhara S (Co-corresponding author), Zhang J, Yuge S, Ando K, Wakayama Y, Sakaue-Sawano A, Miyawaki A, Mochizuki N.
  • Journal Title: Dev. Biol. Volume: 393 Issue: 1 Pages: 10-23
  • DOI: 10.1016/j.ydbio.2014.06.015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. (2014)
  • Author(s): Mort RL, Ford MJ, Sakaue-Sawano A, Lindstrom NO, Casadio A, Douglas AT, Keighren MA, Hohenstein P, Miyawaki A, *Jackson IJ.
  • Journal Title: Cell Cycle Volume: 13 Issue: 17 Pages: 2681-2696
  • DOI: 10.4161/15384101.2015.945381
  • Peer Reviewed
• [Journal Article] A novel cell-cycle indicator, mVenus-p27K-, identifies quiescent cells and visualizes G0-G1 transition. (2014)
  • Author(s): Oki, T., Nishimura, K., Kitaura, J., Togami, K., Maehara, A., Izawa, K., Sakaue-Sawano. A., Niida, A., Miyano, S., Aburatani, H., Kiyonari, H., Miyawaki, A. and Kitamura, T.
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 4012-4012
  • DOI: 10.1038/srep04012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Visualizing developmentally programmed endoreplication in mammals using ubiquitin oscillators (2013)
  • Author(s): Sakaue-Sawano A, Hoshida T, Yo M, Takahashi R, Ohtawa K, Arai T, Takahashi E, Noda S, Miyoshi H, Miyawaki A.
  • Journal Title: Development Volume: 140 Issue: 22 Pages: 4624-4632
  • DOI: 10.1242/dev.099226
  • Peer Reviewed
• [Journal Article] Contrasting Quiescent G0 Phase with Mitotic Cell Cycling in the Mouse Immune System. (2013)
  • Author(s): Tomura M, Sakaue-Sawano A, Mori Y, Takase-Utsugi M, Hata A, Ohtawa K, Kanagawa O, Miyawaki A.
  • Journal Title: Plos One Volume: 8 Issue: 9 Pages: e73801-e73801
  • DOI: 10.1371/journal.pone.0073801
  • Peer Reviewed
• [Presentation] 細胞社会のheterogeneityを視る技術 (2015)
  • Author(s): 阪上-沢野朝子
  • Organizer: 新学術領域ナノメディシン分子科学主催イメージングセミナー
  • Place of Presentation: 東北大学医学部1号館 第１講義室・仙台市・宮城県
  • Year and Date: 2015-09-10
  • Invited
• [Presentation] Observing heterogeneity in cell society (2014)
  • Author(s): Asako Sakaue-Sawano & Atsushi Miyawaki
  • Organizer: 第11回日中合同組織細胞化学セミナー
  • Place of Presentation: 長野県松本市中央公民館（Mウイング文化センター）
  • Year and Date: 2014-09-28 – 2014-09-29
  • Invited
• [Presentation] 細胞社会のheterogeneityを視る技術 (2014)
  • Author(s): 阪上－沢野 朝子
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-26
  • Invited
• [Presentation] Observing heterogeneity in a cell society (2014)
  • Author(s): Asako Sakaue-Sawano & Atsushi Miyawaki
  • Organizer: The 37th Naito Conference
  • Place of Presentation: Hilton Niseko Village, Hokkaido, Japan
  • Year and Date: 2014-07-15 – 2014-07-18
• [Presentation] 細胞動態を4次元で理解するための技術開発 (2014)
  • Author(s): 阪上―沢野 朝子 宮脇 敦史
  • Organizer: 日本顕微鏡学会第70回記念学術講演会
  • Place of Presentation: 幕張メッセ国際会議場
  • Year and Date: 2014-05-11 – 2014-05-13
  • Invited
• [Presentation] 細胞社会を4次元で理解するための技術開発 (2014)
  • Author(s): Asako Sakaue-Sawano & Atsushi Miyawaki
  • Organizer: 日本顕微鏡学会 バイオメディカルニュ－マイクロスコ－プ分科会
  • Place of Presentation: 帝京大学
  • Invited
• [Presentation] Heterogeneity in a cell society (2013)
  • Author(s): Asako Sakaue-Sawano & Atsushi Miyawaki
  • Organizer: RC, Aging, Biology of
  • Place of Presentation: Lucca (Barga), Italy
• [Presentation] 細胞動態を4次元で理解するための技術開発 (2013)
  • Author(s): 阪上―沢野 朝子 宮脇 敦史
  • Organizer: 第86回日本生化学会
  • Place of Presentation: パシフィコ横浜
  • Invited
• [Presentation] Observing heterogeneity in cancer society (2013)
  • Author(s): Asako Sakaue-Sawano & Atsushi Miyawaki
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
  • Invited
• [Remarks] Laboratory for Cell Function Dynamics ＿ Fucci
  • URL: http://cfds.brain.riken.jp/Fucci.html
• [Remarks] Fucci
  • URL: http://cfds.brain.riken.jp/Fucci.html