An object of the present invention is to provide a convenient and low-cost method for enhancing the stability of a DNA aptamer and/or its capacity to bind to a target molecule, and a DNA aptamer obtained by the method. The object is solved by substituting an internal hairpin structure (stem-loop structure) of the DNA aptamer with a structure called mini-hairpin structure and optionally increasing GC pairs in a stem portion of the DNA aptamer.
An object of the present invention is to provide a convenient and low-cost method for enhancing the stability of a DNA aptamer and/or its capacity to bind to a target molecule, and a DNA aptamer obtained by the method.
The object is solved by substituting an internal hairpin structure (stem-loop structure) of the DNA aptamer with a structure called mini-hairpin structure and optionally increasing GC pairs in a stem portion of the DNA aptamer.
The purpose of the present invention is to provide an aptamer to a cancer cell, said aptamer being superior to conventional aptamers in binding ability, specificity and/or stability. To solve this problem, provided is a DNA aptamer binding to a cancer cell, said DNA aptamer containing an artificial base.
C12N 15/115 - Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith
A61K 31/711 - Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
A61K 31/713 - Double-stranded nucleic acids or oligonucleotides
A61K 48/00 - Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseasesGene therapy
The present invention addresses the problem of providing an aptamer for vWF, which is superior in a binding ability, a dissociation rate and/or stability to the conventional nucleic acid aptamers. The present invention can solve the problem by a DNA aptamer which contains an artificial nucleotide and can bond to vWF.
A61P 9/10 - Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
An object of the present invention is to develop and provide a method for efficiently producing a nucleic acid aptamer, particularly, a DNA aptamer, having higher specificity and binding activity against a target substance than those of nucleic acid aptamers obtained by conventional methods. The present invention provides a transcribable or replicable nucleic acid aptamer comprising a natural nucleotide and a non-natural nucleotide having an artificial base-pairable artificial base. The present invention also provides a method for sequencing a non-natural nucleotide-containing single-stranded nucleic acid molecule selected from a single-stranded nucleic acid library.
The present invention addresses the problem of providing a simple and low cost method for improving the stability and/or ability to bind to a target molecule of a DNA aptamer and a DNA aptamer obtained by the method. According to the present invention, the above problem is solved by substituting a hairpin structure (stem-loop structure) within a DNA aptamer by a structure that is called a mini-hairpin structure and arbitrarily increasing GC pairs in the stem part of the DNA aptamer.
An object of the present invention is to develop and provide a method for efficiently producing a nucleic acid aptamer, particularly, a DNA aptamer, having higher specificity and binding activity against a target substance than those of nucleic acid aptamers obtained by conventional methods. The present invention provides a transcribable or replicable nucleic acid aptamer comprising a natural nucleotide and a non-natural nucleotide having an artificial base-pairable artificial base. The present invention also provides a method for sequencing a non-natural nucleotide-containing single-stranded nucleic acid molecule selected from a single-stranded nucleic acid library.
The purpose of the present invention is to develop and provide a method for efficiently producing a nucleic acid aptamer, in particular a DNA aptamer, having a higher specificity and a higher binding ability to a target than nucleic acid aptamers obtained by the conventional methods. Provided is a transcribable or replicable nucleic acid aptamer which comprises a natural nucleotide and a non-natural nucleotide having an artificial base that is capable of pairing with an artificial base. Also provided is a method for determining the base sequence of a non-natural nucleotide-containing single-stranded nucleic acid molecule that is selected from a single-stranded nucleic acid library.
C12N 15/115 - Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith
A61K 31/7115 - Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
A61K 48/00 - Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseasesGene therapy
A61P 9/00 - Drugs for disorders of the cardiovascular system
A61P 43/00 - Drugs for specific purposes, not provided for in groups
C07H 21/02 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
C12N 9/00 - Enzymes, e.g. ligases (6.)ProenzymesCompositions thereofProcesses for preparing, activating, inhibiting, separating, or purifying enzymes
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C40B 40/06 - Libraries containing nucleotides or polynucleotides, or derivatives thereof
Resolution resistance of a single-strand or double-strand nucleic acid fragment containing a base sequence of a functional nucleic acid is strengthened and improved with convenience and at a low cost with respect to a nuclease. In the nucleic acid fragment according to the present invention, at least one 3' end of the single-strand nucleic acid fragment or the double-strand nucleic acid fragment is configured by (A) a nucleic acid domain which is configured by a predetermined number of nucleotides, the number being between two and five, (B) a nucleic acid domain which is configured by a base sequence of gna or gnna (each n herein is, independently, any one of g, t, a, or c, a base analog, or a modified base), and (C) a nucleic acid domain which is configured by a complementary base sequence at the nucleic acid domain of (A), which connect hairpin-type DNAs which are connected in sequence from a 5' end to the 3'end. At least one of the two nucleotides is modified from at least the one 3' end of the single-strand nucleic acid fragment or the double-strand nucleic acid fragment.
In order to develop and provide a method for conveniently and efficiently producing a nucleic acid aptamer, in particular a DNA aptamer, having excellent bonding and specificity properties with a target substance, a production method for a nucleic acid aptamer comprises: a complex formation step in which a complex of a single-stranded nucleic acid and a target substance is formed by mixing, in a solution, a single-stranded nucleic acid library and the target substance; an immobilization step in which a solid support and the solution after the above-mentioned step are mixed, and the complex is immobilized on the solid support via the target substance and/or bonding elements adhered to the solid support; a recovery step in which the complex, immobilized on the solid support, is recovered from the solution; an amplification step in which the single-stranded nucleic acid is recovered from the complex, and then amplified by means of a nucleic acid amplification method; and a single-stranded nucleic acid preparation step in which double-stranded nucleic acid, obtained in the amplification step, is made to be single stranded, and then an in-molecule three-dimensional structure is formed.
C12N 15/115 - Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith
A61K 31/7088 - Compounds having three or more nucleosides or nucleotides
A61K 31/711 - Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
A61K 31/713 - Double-stranded nucleic acids or oligonucleotides
A61P 43/00 - Drugs for specific purposes, not provided for in groups
The present invention relates to novel unnatural fluorescent nucleic acid bases, that is, a purine base, a 1-deazapurine base, and a 1,7-deazapurine base each having a functional group which consists of two or more heterocyclic moieties linked together, at the 6-position thereof (the 6-position of purine ring). The present invention also relates to a compound containing the unnatural base, a derivative thereof, and a nucleic acid containing a nucleotide having the unnatural base. The present invention also relates to a method of preparing the nucleic acid. The unnatural base of the present invention has excellent fluorescence characteristics and also has excellent properties as a universal base.
C07H 19/00 - Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radicalNucleosidesMononucleotidesAnhydro derivatives thereof
The present invention provides a double-stranded nucleic acid in which at least one nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability. The present invention also provides a method of hybridizing a first nucleic acid strand with a second nucleic acid strand, wherein the first nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability, and a method of applying the nucleic acid to SNP detection, a DNA chip, DNA/RNA computing, or an in vitro translation system. The present invention provides a method of introducing an unnatural base into a nucleic acid strand and thereby controlling the thermodynamic stability in hybridization of the nucleic acid strand.
C07H 21/02 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
Disclosed is a simple and low-cost method for strengthening and improving the nuclease degradation resistance of single-chain or two-chain nucleic acid fragments containing a base sequence of functional nucleic acids. Specifically, hairpin DNA is attached to at least one of the ends of the single-chain nucleic acid fragment or two-chain nucleic acid fragment. This hairpin DNA is formed from a nucleic acid region (A) formed from 2 - 5 random nucleotides, a nucleic acid region (B) formed from a base sequence of gna or gnna (wherein each n is, independently, any of g, t, a, or c, a base analog or a modified base), and a nucleic acid region (C) formed from a complementary base sequence for nucleic acid region (A) such that these regions are attached sequentially from the 5' end side toward the 3' end side.
C12N 15/113 - Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides
A61K 31/7088 - Compounds having three or more nucleosides or nucleotides
A61K 31/7105 - Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
A61K 31/711 - Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
A61K 48/00 - Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseasesGene therapy
A61P 43/00 - Drugs for specific purposes, not provided for in groups
Disclosed is a nucleic acid base analog with quenching characteristics and fluorescence and application thereof. Specifically disclosed is a quencher characterized by having a 2-nitropyrrole structure shown by formula (I). [In formula (I), R1 and R2 are groups selected independently from a group formed from ribose and deoxyribose; hydrogen, hydroxyl groups, SH groups, and halogens; optionally substituted C2-10 alkyl groups, alkenyl groups, and alkynyl groups; one or a plurality of 5-member heterocycles, one or a plurality of 6-member heterocycles, one or a plurality of polycyclic heterocycles and one or a plurality of aromatic rings, containing nitrogen atoms or sulfur atoms; sugars, sugar chains, amino acids, and peptides; and florescent molecules bound through linkers.]
G01N 33/542 - ImmunoassayBiospecific binding assayMaterials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
15.
ARTIFICIAL BASE PAIR CAPABLE OF FORMING SPECIFIC BASE PAIR
Disclosed is a double-stranded nucleic acid wherein at least one of the nucleic acids has an artificial base capable of forming a self-complementary base pair or an artificial base capable of forming a base pair of a heat stability at the same level with any natural base. Also disclosed are a method for hybridizing a first nucleic acid, which has an artificial base capable of forming a self-complementary base pair or an artificial base capable of forming a base pair of a heat stability at the same level with any natural base, with a second nucleic acid, and a method for using said first nucleic acid in detecting an SNP or applying the same to a DNA chip, DNA/RNA computing or an in vitro translation system. Also disclosed is a method for manipulating the thermodynamic stability in the hybridization of a nucleic acid by introducing an artificial base into said nucleic acid.
C07H 21/02 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
C07H 19/23 - Heterocyclic radicals containing two or more heterocyclic rings condensed among themselves or condensed with a common carbocyclic ring system, not provided for in groups
C07H 19/24 - Heterocyclic radicals containing oxygen or sulfur as ring hetero atom
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
C12M 1/00 - Apparatus for enzymology or microbiology
Disclosed is a novel artificial fluorescent nucleic acid base having a functional group, which consists of two or more heterocyclic molecules being polycondensed together, at the 6-position (the 6-position of purine ring) of a purine base, a 2-deazapurine base or a 2,7-deazapurine base. Also disclosed are a compound containing said artificial base, a derivative of the same, and a nucleic acid having a nucleotide, which contains said artificial base, integrated therein. Moreover, disclosed is a method for preparing said nucleic acid. The aforesaid artificial base has excellent fluorescent characteristics and shows excellent properties as a universal base.
C07H 19/23 - Heterocyclic radicals containing two or more heterocyclic rings condensed among themselves or condensed with a common carbocyclic ring system, not provided for in groups
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
C12N 15/00 - Mutation or genetic engineeringDNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purificationUse of hosts therefor
17.
DNA capable of being amplified by PCR with high selectivity and high efficiency
The present invention relates to unnatural base pairs of Ds (a 7-(2 thienyl)-3H-imidazo[4,5-b]pyridine-3-yl group) and a Pa derivative (a 2-nitro-1H-pyrrole-1-yl group bearing a substituent having a π-electron system attached at position 4) that can be replicated with high selectivity/high efficiency, and methods for replicating nucleic acids containing the unnatural base pairs. The present invention also relates to methods for incorporating an unnatural base bearing a functional substituent attached thereto into DNA by a nucleic acid replication reaction. The present invention also relates to methods for replicating and selectively collecting a nucleic acid containing an unnatural base pair from a nucleic acid pool. The present invention also relates to methods for determining a sequence of natural bases in the proximity of an unnatural base in DNA for achieving highly efficient and highly selective replication of a nucleic acid containing the unnatural base.
C07H 21/02 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
Disclosed are: an artificial base pair Ds-(Pa derivative) [wherein Ds represents a 5-amino-7-(2-thienyl)-3H-imidazo[4,5-b]pyridin-3-yl group; and the Pa derivative is a 2-nitro-1H-pyrrol-1-yl group having a substituent bearing a ꧀-electron system bound to position-4 thereof], which can be replicated with high selectivity and high efficiency; a method for amplifying a nucleic acid containing the artificial base pair; a method for introducing an artificial base having a functional substituent bound thereto into DNA through a nucleic acid amplification reaction; a method for replicating a nucleic acid containing an artificial base pair from a nucleic acid pool and collecting the replicated nucleic acid selectively; and a method for determining a naturally occurring nucleotide sequence located adjacent to an artificial base in DNA for the purpose of achieving the highly efficient and highly selective replication of a nucleic acid containing an artificial base.
A61K 31/7115 - Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
The present invention addresses the problem of providing an aptamer for vWF, which is superior in a binding ability, a dissociation rate and/or stability to the conventional nucleic acid aptamers. The present invention can solve the problem by a DNA aptamer which contains an artificial nucleotide and can bond to vWF.
A61P 9/10 - Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
C12N 15/115 - Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
Disclosed are: an artificial base pair Ds-(Pa derivative) [wherein Ds represents a 5-amino-7-(2-thienyl)-3H-imidazo[4,5-b] pyridin-3-yl group; and the Pa derivative is a 2-nitro-1H-pyrrol-1-yl group having a substituent bearing a .pi.-electron system bound to position-4 thereof], which can be replicated with high selectivity and high efficiency; a method for amplifying a nucleic acid containing the artificial base pair; a method for introducing an artificial base having a functional substituent bound thereto into DNA through a nucleic acid amplification reaction; a method for replicating a nucleic acid containing an artificial base pair from a nucleic acid pool and collecting the replicated nucleic acid selectively; and a method for determining a naturally occurring nucleotide sequence located adjacent to an artificial base in DNA for the purpose of achieving the highly efficient and highly selective replication of a nucleic acid containing an artificial base.
A61K 31/7115 - Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
patents
