Abstract: It is an object of the present invention to provide an RpBphP1 mutant and an RpPpsR2 mutant that have been improved so that their binding and dissociation can be reliably controlled by light irradiation with a near-infrared light and blocking thereof. More specifically, the present invention relates to a protein set consisting of two proteins that bind to each other under near-infrared light irradiation, the protein set being a set of an RpBphP1 mutant and a wild-type RpPpsR2, a set of a wild-type RpBphP1 and an RpPpsR2 mutant, or a set of an RpBphP1 mutant and an RpPpsR2 mutant, wherein the binding of the protein set is weaker under the dark and/or is stronger under near-infrared light irradiation, than the binding of a wild-type RpBphP1 and a wild-type RpPpsR2.

Abstract: The present invention provides a set of Affibody and photoreceptor protein, wherein the Affibody and the photoreceptor protein form a complex in a manner dependent on light at a wavelength of 600 to 750 nm.

Abstract: 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.

Abstract: It is an object of the present invention to provide a method of reliably chemogenetically controlling the activity of a protein, and to provide a protein used in the present method. Specifically, the present invention relates to a plurality of protein fragments, which recover the activity of the protein when the plurality of protein fragments assemble, and in which each of the plurality of protein fragments is tagged with a degradation domain (DD) sequence. In addition, the present invention also relates to a method of regulating the activity of a protein in a cell, comprising: a step of introducing into a cell, a fusion of a DD sequence, and each fragment of a protein that has been split into fragments, such that the fragments recover the activity of the protein when they assemble; and a step of introducing a DD sequence-specific stabilizing factor into the cell.

Abstract: 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.

Abstract: The present invention provides a set of two polypeptides of a split Cpf1 protein, wherein the two polypeptides are a N-terminal side fragment of a Cpf1 protein and a C-terminal side fragment of the Cpf1 protein.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: Probe 1 for detection and quantification of nitric oxide, which comprises two subunits 21 and 22 of soluble guanylate cyclase 2 and cGMP-visualization probes 3 respectively connected with each subunit, wherein the cGMP-visualization probe generates signal upon recognizing guanosine 3?,5?-cyclic monophospate.

Abstract: 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.

Abstract: 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.

Abstract: This invention provides a genetically modified marine luciferase such as Gaussia luciferase, which has high bioluminescence intensity, and has high bioluminescence stability and/or red-shifted wavelength. Specifically disclosed is a luciferase variant with improved optical property obtained by replacing at least one amino acid residue among the amino acid sequence of a marine luciferase at positions corresponding to positions 89 to 118 in the amino acid sequence of Gaussia luciferase (GLuc), wherein an amino acid residue at a position corresponding to at least one selected from positions 89, 90, 95, 97, 100, 108, 112, 115, and 118 in the amino acid sequence of GLuc is replaced by way of conservative amino acid replacement. The above-mentioned replacement in a marine luciferase improves enzymatic activity of the luciferase. Also disclosed is a bioluminescent probe having an improved optical property, which is produced using the luciferase variant of the present invention.

Abstract: Probe 1 for detection and quantification of nitric oxide, which comprises two subunits 21 and 22 of soluble guanylate cyclase 2 and cGMP-visualization probes 3 respectively connected with each subunit, wherein the cGMP-visualization probe generates signal upon recognizing guanosine 3?,5?-cyclic monophospate.

Abstract: 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).

Abstract: This application aims to provide a single molecule-format probe for detecting a target-specific ligand easily and accurately as an index of the presence or absence of a signal. The invention is 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, and an N-terminal polypeptide (N-LE) and a C-terminal polypeptide (C-LE) of a luminescent enzyme (LE), which are linked to each end of the ligand-binding molecule, respectively, wherein the N-LE and the C-LE self-complement to generate a luminescent signal only upon binding of the ligand to the ligand-binding molecule. The ligand-binding molecule is a fusion molecule carrying a ligand-binding domain (LBD) and an LBD-interacting domain or peptide that interacts with the LBD upon binding of the ligand to the LBD.

Abstract: 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.