Japan Science and Technology Agency

Japan Advanced Institute of Science and Technology

Nicca Chemical Co., Ltd.

Lsi Medience Corporation

Toray Industries, Inc.

Kenzo Fujimoto

Yoshinaga Yoshimura

Masayuki Ogino

Hiroshi Terasaki

Mitsunobu Shimadzu

Shigetaka Nakamura

Takashi Sakamoto

Yuya Tanaka

Yoji Ueda

Fumio Nakamura

Taisuke Hirano

Hirotake Wakamatsu

Shinzi Ogasawara

Yukari Nitta

Shinya Toba

Yuta Taya

Tsunetada Konno

Masahiko Takamura

Isao Ishimaru

Ayako Koto

Takahiro Inatsugi

Nobutake Fugono

Akira Yanagihara

Provided are: a photocoupling method that overcomes the problem of the stagnation of photocoupling with a target nucleotide using a probe containing a photo-responsive nucleotide, and that improves the photocoupling efficiency; and a photocoupling kit. The photocoupling method is characterized by hybridizing a target site present in a nucleic acid sample with a first probe having a sequence complementary to the target site and containing a photo-responsive nucleotide, in a reaction solution, and carrying out photocoupling by photo-irradiation, wherein self-assembly caused by the photo-responsive nucleotide contained in the first probe is suppressed. The photocoupling kit is characterized by comprising a first probe having a sequence complementary to a target site present in a nucleic acid sample, and containing a photo-responsive nucleotide; and a second probe being highly complementary to the first probe.

Photocoupling method using probe containing photoresponsive nucleic acids

Provided are a compound represented by formula I that can be used in nucleic acid photoreaction techniques, and a photoreactive crosslinking agent comprising the compound. 

Photoresponsive nucleotide analog capable of photocrosslinking in visible light region

Provided are a compound represented by the following formula I; a novel photoreactive compound that can be used in nucleic acid photoreaction techniques by a photoreactive crosslinking agent comprising the compound; and a photoreactive crosslinking agent in which the photoreactive compound is used. 

Provided is a means for preventing the inactivation of a photoresponsive nucleic acid probe by suppressing the formation of a photocrosslink between a modified nucleotide having a structure corresponding to the monomer of Formula (II) or an amino acid analogue of a nucleotide having a structure corresponding to the monomer of Formula (III) and a modified nucleotide having a structure corresponding to the monomer of Formula (I), wherein the modified nucleotide replaces at least one constituent nucleotide which is the photocrosslinkable 1-thyminyl or 1-uracilyl, by substituting at least one constituent nucleotide which is the photocrosslinkable 1-thyminyl or 1-uracilyl with a modified nucleotide having a structure corresponding to the monomer of Formula (I).

Method of suppressing formation of photocrosslink, and photoreactive nucleic acid in which auto-crosslink formation is suppressed

The present invention provides a new photoreactive compound which can be used in technologies for photoreactions of nucleic acid, and also provides a photoreactive crosslinking agent comprising the above photoreactive compound. A photoreactive compound represented by the following formula I can be used. 

Photoresponsive nucleotide analogue having photocrosslinking ability

Provided is a method for rapidly and easily detecting a mutated nucleic acid, which is contained in a small amount in a nucleic acid sample together with wild-type nucleic acids, with high specificity and high sensitivity. In the method of the present invention, amplification of a detection region comprising a target site by a nucleic acid amplification method is inhibited, by the steps of allowing a nucleic acid having a target site to coexist with a clamp probe comprising a photo-crosslinking nucleic acid and having a sequence complementary to the target site, and photo-crosslinking the nucleic acid having the target site with the clamp probe by photo-irradiation.

Inhibition method of nucleic acid amplification by photoirradiation and method of selective nucleic acid amplification with high sensitivity

A method of detecting a target nucleic acid by sandwich hybridization using a detection probe that hybridizes with the target nucleic acid, and a capture probe immobilized on a support wherein at least one nucleic acid base in the nucleic acid molecule(s) of the detection probe and/or capture probe with a photoreactive group includes irradiating, with light, a complex formed by hybridization between the target nucleic acid and the detection probe and/or capture probe to allow formation of a covalent bond(s) between the photoreactive group(s) and a nucleic acid base(s) in the target nucleic acid.

Method of detecting target nucleic acid

The present invention provides a manufacturing method that can easily manufacture a compound known as photoresponsive (photocoupling) nucleic acids at high yield in a shorter period of time than that of the conventional technology. The present invention relates to a method of manufacturing a photoresponsive nucleic acid which includes a step of reacting a nucleic acid having groups represented by the Formula I, the Formula III, the Formula IV, or the Formula V and a compound represented by the Formula II, or reacting a nucleic acid having groups represented by the Formula VI, the Formula VIII, the Formula IX, or the Formula X and a compound represented by the Formula VII by heating them by microwaves in the presence of a metal catalyst, a basic substance, and a solvent.

Photoresponsive nucleic acid manufacturing method

The present invention provides a method for detecting methylcytosine in DNA rapidly, conveniently, and with high sensitivity. The present invention relates to a method for detecting methylcytosine by using a methylcytosine photocoupling agent (a photoresponsive probe) consisting of nucleic acids having a group represented by the Formula (I), (II), (III) or (IV) as a base moiety.

Method for detection of methylcytosine using photoresponsive probe

The present invention provides a photoreactive crosslinking agent that is capable of crosslinking a sequence which cannot be photo-crosslinked by psoralen, and is capable of photo-crosslinking using a light having a longer wavelength, as compared with psoralen. The present invention also provides a compound having a group represented by formula (I) coupled with a group represented by formula (II).

Light-responsive artificial nucleotide having photo-crosslinking ability

Disclosed is a method for the purification and collection of a nucleic acid comprising a specific nucleotide sequence, which can be carried out within an extremely short period and can achieve both high sequence-specificity and a high collection rate. Specifically disclosed is a method for the purification of a target nucleic acid comprising a specific nucleotide sequence and contained in a nucleic acid mixture. The method comprises the steps of: hybridizing a photo-ligating nucleic acid having a group represented by formula (I) as abase moiety with the target nucleic acid to form a hybrid; irradiating the hybrid of the photo-ligating nucleic acid and the target nucleic acid with light to cause the photo-ligation of the hybrid; removing any un-photo-ligated nucleic acid by washing; and irradiating the hybrid of the photo-ligating nucleic acid and the target nucleic acid with light to cause the photo-cleavage of the hybrid.

Sequence-specific nucleic acid purification method manner

It is intended to provide a method for detecting a target nucleic acid with a specific base sequence existing in a sample mixture with high specificity and sensitivity by utilizing the formation of a hybrid with a complementary strand as a detection principle, and a nucleic acid for the detection. The invention relates to a set of nucleic acids for detecting a target nucleic acid with a specific base sequence comprising a photoligating nucleic acid composed of a nucleic acid (with the proviso that in the nucleic acid, a nucleic acid and a peptide nucleic acid are included) having a group represented by the formula I, II, III, IV or V described in claims at the 5' end or 3&#x2032; end as a base moiety, and a photoligated nucleic acid having a base with a carbon-carbon double bond at the 3&#x2032; end or 5&#x2032; end as a base moiety capable of photoligating to the photoligating nucleic acid, wherein either of the photoligating nucleic acid and the photoligated nucleic acid has a labeling moiety and the other remaining nucleic acid is immobilized on a substrate, and the method for detecting a target nucleic acid with a specific base sequence by using the set of nucleic acids.

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Nomi, Japan

Kenzo Fujimoto