| Project/Area Number |
17017039
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
|
| Allocation Type | Single-year Grants |
|---|
| Review Section |
Biological Sciences
|
|---|
| Research Institution | The Institute of Physical and Chemical Research |
|---|
Principal Investigator |
MIYAWAKI Atsushi The Institute of Physical and Chemical Research, 細胞機能探索技術開発チーム, チームリーダー (80251445)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIZUNO Hideaki 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 専門職研究員 (80301779)
SHIMOZONO Satoshi 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 研究員 (40391982)
FUKANO Takashi 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 研究員 (80373364)
TSUTSUI Hidekazu 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 客員研究員 (30392038)
NAGAI Takeharu 北海道大学, 電子科学研究所, 教授 (20311350)
|
|---|
| Research Collaborator |
IBATA Keiji 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 研究員
MIYAUCHI Takayuki 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 研究員 (00392142)
VETRIVEL Lakshmanan 独立行政法人理化学研究所, 細胞機能探索技術開発チーム, 研究員
|
|---|
| Project Period (FY) |
2005 – 2009
|
| Project Status |
Completed (Fiscal Year 2009)
|
| Budget Amount *help |
¥54,900,000 (Direct Cost: ¥54,900,000)
Fiscal Year 2009: ¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 2008: ¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 2007: ¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2006: ¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 2005: ¥7,100,000 (Direct Cost: ¥7,100,000)
|
|---|
| Keywords | 蛍光蛋白質 / 細胞周期 / レシオイメージング / 膜電位 / 蛍光イメージング / 神経前駆細胞 / 蛍光タンパク質 / バイオイメージング |
|---|
| Research Abstract |
We harnessed the regulation of cell-cycle-dependent ubiquitination to develop a genetically encoded indicator for cell-cycle progression, Fucci. This technology permits us to visualize the cell-cycle behavior of individual cells within complex tissues of experimental animals, such as mice and fish. We used two new coral fluorescent proteins as FRET donor and acceptor to develop a voltage sensor, Mermaid. This technology allows for direct visualization of electrical activities in excitable cells. We developed a far-red fluorescent protein endowed with a large Stokes shift, Keima. We show the usefulness of Keima for dual-color fluorescence imaging technologies, such as fluorescence cross-correlation spectroscopy and two-photon excitation microscopy.
The structural basis for the photochromism in the fluorescent protein Dronpa is poorly understood. We performed NMR analyses of Dronpa in solution at ambient temperatures to find structural flexibility of the protein in the dark state. We have also developed numerous mutants of Dronpa, which change between bright and dark states with different speeds. These mutants were used to improve performance of super-resolution microscopy. KikGR is a fluorescent protein engineered to display green-to-red photoconvertibility that is induced by irradiation with violet light. Through crystallographic studies on both green and red states, we obtained evidence that the β-elimination reaction governing the green-to-red photoconversion follows an E1 (unimolecular) mechanism. Using KikGR as an optical highlighter, we demonstrate that diffusion barrier across the nuclear envelope is less restrictive during nuclear reassembly.
|
