National Institute of Advanced Industrial Science and Technology

The University of Tokyo

Japan Science and Technology Corporation

Japan Science and Technology Agency

Keio University

Moritoshi Sato

Yoshio Umezawa

Hiroaki Tao

Takeaki Ozawa

Sung-Bae Kim

Koji Suzuki

Takahiro Nakajima

Makoto Takeda

Daniel Citterio

Kenzaburo Tani

Yuta Nihongaki

Ryo Nishihara

SungBae Kim

Maino Tahara

The purpose of the present invention is to provide a set of two polypeptides for use in light-dependent genetic recombination in which the N-terminal side fragment and the C-terminal side fragment of a Cre protein having an amino acid sequence of SEQ ID NO: 1 respectively bind to two proteins light-dependently forming a dimer.

Polypeptide set to be used in light dependent gene recombination

The purpose of the present invention is to develop a virus vector, the activity of which is rendered controllable. A virus protein gene derived from an RNA virus is provided in which a gene encoding an optical switch protein is inserted into a foreign gene introducible region of the virus protein so as to enable expression of the gene. By means of this virus vector, it is possible to control, with irradiation of light, enzyme activity of the virus protein and virus vector activity based thereon.

Optically controlled virus protein, gene thereof, and virus vector containing said gene

The present invention provides, for example, a set of two polypeptides exhibiting the nuclease activity with dependence on light or in the presence of a drug, in which an N-terminal side fragment and a C-terminal side fragment of a Cas9 protein are bound to each of two polypeptides which form a dimer with dependence on light or in the presence of a drug.

Set of polypeptides exhibiting nuclease activity or nickase activity with dependence on light or in presence of drug or suppressing or activating expression of target gene

There is provided a series of coelenterazine (CTZ) derivatives as a substrate with high luminescence intensity, which is optimal for maximum luminescence wavelengths at both 400 nm (blue-shifted RLuc luminescence system) and 500 nm (ALuc luminescence system) for bioassays which is more sensitive than known techniques. The novel CTZ derivatives are compounds in which a specific position(s) of the CTZ is/are substituted with a specific substituent(s) as shown, for example, by the Formula [I], and has a higher luminescence intensity than known CTZ derivatives in blue-shifted RLuc luminescence system or ALuc luminescence system.

Coelenterazine compounds and uses thereof

Probefor detection and quantification of nitric oxide, which comprises two subunitsandof soluble guanylate cyclaseand cGMP-visualization probesrespectively connected with each subunit, wherein the cGMP-visualization probe generates signal upon recognizing guanosine 3',5&#x2032;-cyclic monophospate.

Probe for detection and quantification of nitric oxide, and method for detecting and quantifying nitric oxide using the same

A single-chain probe of the present invention for detecting a ligand, comprises: a ligand binding protein for binding the ligand; a recognition protein for recognizing that the ligand is bound by the ligand binding protein; and C- and N-terminal fragments, generated by dissecting an enzyme, between the ligand binding protein and the recognition protein, wherein a carboxy terminal end of the C-terminal fragment is located upstream of an amino terminal end of the N-terminal fragment, and the C- and N-terminal fragments vary the enzyme activity via complementation in case where the recognition protein recognizes that the ligand is bound by the ligand binding protein. This makes it possible to achieve detection of a target protein-specific ligand using the single chain with a high efficiency.

Single-molecule-format probe and utilization thereof

The claimed invention comprises a single molecule-format bioluminescent probe for detecting a target-specific ligand in a living cell, which comprises, a ligand-binding molecule of which conformation is changed upon binding to the ligand, wherein the ligand-binding molecule comprises a ligand-binding domain (LBD) of a nuclear receptor and an LBD-interacting domain that is a co-activator peptide of said nuclear receptor, and an N-terminal polypeptide and a C-terminal polypeptide of a click beetle luciferase (N-CBLuc and C-CBLuc), which flank each end of the ligand-binding molecule, respectively, wherein the N-CBLuc and the C-CBLuc self-complement to generate a luminescent signal only upon binding of the ligand to the ligand-binding molecule.

Single molecule-format bioluminescent probe

The present invention provides an 'in vivo and in vitro real-time bioluminescence imaging means,' which can transmit a detection signal promptly in response to an external signal, while taking advantage of a single-molecule-format luminescent probe as a bioluminescent means. The present invention is characterized by using, as a single-molecule-format luminescent probe utilizing the increase and decrease of a second messenger level as an index, a fusion protein including a single-chain protein containing a second messenger recognition protein and optionally a peptide which is capable of binding with the protein, and linked respectively to the N-terminus and the C-terminus thereto, an N-terminal fragment (N-LE) and a C-terminal fragment (C-LE) generated by dissecting a luminescent enzyme (LE). A single-molecule-format luminescent probe could be provided, which makes it possible to observe light emission (extinction) in vivo as well as in vitro, as a result of a conformational change of the second messenger recognition protein induced by the binding (unbinding) with a second messenger, and the subsequent association or dissociation between the N-LE and the C-LE flanked at both termini of the recognition protein.

Single molecule-format real-time bioluminescence imaging probe

The present invention provides probes for detection and quantification of a lipid second messenger, which comprises: a polypeptide specifically bound to the lipid second messenger; two chromophores respectively having different fluorescence wavelengths, wherein each of the chromophores is linked to each end of the polypeptide through a rigid linker sequence; and a membrane localization sequence linked to one of the chromophores through a rigid linker sequence. According to the present invention, it is now possible to quantitatively detect when and in which site of a living cell the lipid second messengers are produced.

Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same

As a versatile method of detecting and assaying intracellular protein phosphorylation and dephosphorylation that enables nondestructive monitoring as well as spatial and temporal analysis for living cells, animal bodies, plant bodies and the like, a probe for imaging protein phosphorylation and dephosphorylation, which comprises a tandem fusion unit composed of a substrate domain that contains a phosphorylation and dephosphorylation site, a linker sequence and a phosphorylation recognition domain, interposed between a donor chromophore and an acceptor chromophore that cause fluorescence resonance energy transfer, is used.

Probe for visualizing phosphorylation/dephosphorylation of protein and method of detecting and quantifying phosphorylation/dephosphorylation of protein

As a cGMP-visualizing probe capable of detecting and quantifying cGMP easily and accurately even in vivo and for a method of detecting and quantifying cGMP by using the same, a cGMP-visualizing probe comprising a polypeptide, which binds specifically to cGMP, and two chromophores having different fluorescence wavelengths each linked respectively to the two terminals of said polypeptide is provided.

Growing community of inventors

Tokyo, Japan

Moritoshi Sato