96 results about "Minor groove" patented technology

Modified oligonucleotides are provided containing bases selected from unsubstituted and 3-substituted pyrazolo[3,4-d]pyrimidines and 5-substituted pyrimidines, and optionally have attached minor groove binders and reporter groups.

Compositions and methods for modulating the activity of RNA and DNA are disclosed. In accordance with preferred embodiments, antisense compositions are prepared comprising targeting and reactive portions. Reactive portions which act, alternatively, through phosphorodiester bond cleavage, through backbone sugar bond cleavage or through base modification are preferrably employed. Groups which improve the pharmacodynamic and pharmacokinetic properties of the oligonucleotides are also useful in accordance with certain embodiments of this invention. Delivery of the reactive or non-reactive functionalities into the minor groove formed by the hybridization of the composition with the target RNA is also preferrably accomplished. Therapeutics, diagnostics and research methods and also disclosed. Synthetic nucleosides and nucleoside fragments are also provided useful for elaboration of oligonucleotides and oligonucleotide analogs for such purposes.

Modified oligonucleotides are provided containing bases selected from unsubstituted and 3-substituted pyrazolo[3,4-d]pyrimidines and 5-substituted pyrimidines, and optionally have attached minor groove binders and reporter groups.

The invention provides methods and kits for detecting and / or quantifying nucleic acid sequences of interest by using pairs of probes containing donor-acceptor moieties that when hybridized on a target polynucleotide, with one of the probes being hybridized to a sequence of interest in the target polynucleotide, places the donor-acceptor moieties in sufficiently close proximity such that a detectable signal is generated. Methods of the invention are particularly useful for genotyping analysis and gene expression profiling. Methods of the invention are easily adaptable to arrays and automation. The invention also provides probes with enhanced hybridization efficiency and / or specificity comprising a probe portion having a spacer element and / or a minor groove binder molecule, and methods and kits for using these probes.

At least one bioactive agent is locally delivered to a location where a stent is implanted within a lumen in a patient's body. The bioactive agent includes a: DNA minor groove binder (such as CC-1065 or Duocarmycin); apocynin; RGD peptide (such as RGDfV); stilbene compound (such as resveratrol); camptothecin; des-aspartate angiotensin I; or ADF; or an analog or derivative thereof; or a combination or blend thereof with at least one other bioactive agent. The bioactive agent is generally locally delivered, such as by elution from the stent. The compounds and methods are of particular benefit for treating or preventing atherosclerosis, stenosis, restenosis, smooth muscle cell proliferation, occlusive disease, or other abnormal lumenal cellular proliferation condition.

Minor groove binder-oligonucleotide probes are provided along with methods for their use wherein the probes have an attached fluorophore which, in an unhybridized form exhibits very low background signal.

The present invention relates to (i) conjugates comprising two DNA alkylating subunits linked by a moiety fitting to the minor groove of the DNA, (ii) to their preparation and (iii) to their use in cancer therapy. The alkylating subunits are especially cytotoxic under hypoxic conditions found in cancer cells. The compounds of the present invention and compositions thereof are useful in the treatment of cancer in a mammal, both alone or in a combination with other anti-cancer agents (e.g. checkpoint abrogators) and / or radiation. They may also be used as cytotoxic units for gene-directed enzyme-prodrug therapy (GDEPT) and antibody-directed enzyme-prodrug therapy (ADEPT).The present invention provides the compounds of Formula (I), Formula (II) and Formula (III):for treating cancer in a mammal.

The present invention provides a pneumatic tire which can obtain a high wet drainage performance, roadholding ability, and wear-resistant property without sacrificing other capabilities A directional pattern is formed by a circumferential wide major groove 14, a first narrow circumferential minor groove 16, a second narrow circumferential minor groove 18, a first transverse groove 20, a second transverse groove 22, and a third transverse groove 24. The circumferential wide major groove 14 is provided in a center region in a tire width direction of a tread 12. The first narrow circumferential minor groove 16 is arranged on the outside in a tire axis direction of the circumferential wide major groove 14. The second narrow circumferential minor groove 18 is arranged on the outside in the tire axis direction of the first narrow circumferential minor groove 16. The first transverse groove 20 is extended from a tread end 12E and coupled to the circumferential wide major groove 14. The second transverse groove 22 is arranged between the first transverse grooves 20, and the second transverse groove 22 is extended from the tread end 12E and terminated while not coupled to the circumferential wide major groove 14. The third transverse groove 24 is arranged between the first transverse groove 20 and the second transverse groove 22, and the third transverse groove 24 is extended from the tread end 12E and terminated between the first narrow circumferential minor groove 16 and the second narrow circumferential minor groove 18. Therefore, in wet road surface driving, the water between the pneumatic tire and a road surface flows efficiently into each groove to obtain high wet performance. Because the high wet performance is obtained while an increase in negative ratio is suppressed, a wheel tread area of a block is secured to improve a wear-resistant property. A circumferential length of a second block 28 becomes a substantial half of a first block 26, and circumferential lengths of a stepping-on side third block 30 and a kickout side third block 32 become a substantial half of the second block 28. Therefore, a wet drainage property can be improved on both sides of the tread while block rigidity is increased on a tread central region to improve traction performance, brake performance, and cornering performance. Furthermore, when the pneumatic tire of the invention is used in a front wheel, handling performance is improved by increasing the block rigidity in a central region in the tire axis direction. Widths and depths of the first narrow circumferential minor groove 16 and second narrow circumferential minor groove 18 are decreased from a kickout side toward a stepping-on side of the block in a range where the block is zoned by the first and second narrow circumferential minor grooves 16 and 18. Therefore, the block rigidity is increased on the stepping-on side of the blocks located on both sides of the first and second narrow circumferential minor grooves 16 and 18, and thereby the traction performance, brake performance, and cornering performance are improved.

The present invention provides a negatively charged minor groove binding compound, oligonucleotide conjugates comprising the same, and methods for using the same. The negatively charged minor groove binding compounds of the present invention comprises an acidic moiety that is capable of being ionized under physiological conditions. In particular, the negatively charged minor groove binding compound of the present invention comprises a binding moiety that binds preferentially into a minor groove of a double, triple or higher stranded DNA, RNA, PNA or hybrids thereof. The binding moiety comprises at least one aryl moiety and an acidic moiety which is covalently attached to a phenyl portion of the aryl moiety or to a heteroatom of a heteroaryl portion of the aryl moiety.

The present invention provides a new class of solids supports for synthesis of modified oligomers of nucleic acids, and nucleic acid probes that have a format expediently synthesized on the new supports. Exemplary solid supports include at least one quencher bound through a linker to the solid support. Various exemplary embodiments include a moiety that stabilizes a duplex, triplex or higher order aggregation (e.g., hybridization) of nucleic acids of which the oligomer of the invention is a component. Other components of the solid support include moieties that stabilize aggregations of nucleic acids, e.g., intercalators, minor groove binding moieties, bases modified with a stabilizing moiety (e.g., alkynyl moieties, and fluoroalkyl moieties), and conformational stabilizing moieties, such as those described in commonly owned U.S. Patent Application Publication No. 2007 / 0059752.

Oligonucleotide probes containing two labels are provided and are useful in hybridization assays. The probes can also contain a minor groove binding group.

The invention discloses a primer pair for specific amplification of EGFR gene 20 exon T790M and C797S loci. The sequence of the primer pair is shown in SEQ ID No:1-2. The invention further discloses a probe set for specific detection of the T790M and C797S loci. The probe set comprises a wild type probe and a mutant type probe, and the sequences are shown in SEQ ID No:3-7. The invention further discloses a reagent kit comprising the primer pair and the probe set, and a method for adopting the primer pair and the detection set for detecting EGFR gene 20 exon T790M and C797S mutation. By designing the specific primers and probes, the 3' end of the probes is connected with a minor groove binder and a non-fluorescent quenching group, the interference of background signals is lowered, hybridization of the probes and a template is stabilized, a Tm value of the probes is increased, and therefore the mutation detection sensitivity is improved.

The invention relates to a kit for simultaneously detecting statin metabolizing gene multisite mutations. The kit includes primer pairs and probe pairs for detecting APOE, SLCO1B1, CETP, ABCB1 and MTHFR gene sites. MGB (minor groove binder) probes and a real-time fluorescent PCR technique are applied in the kit. The kit has advantages of capability of being time saving and convenient, high sensitivity, capability of allowing positive and negative coincidence rates of a sample to be 99.5% or above, and the like. The kit is mainly used for personalized medication assisted diagnosis of statins such as simvastatin, atorvastatin and pravastatin.

The invention discloses a human K-ras (K-rat sarcoma) gene mutation parting type fluorescent quantitation PCR (polymerase chain reaction) detecting reagent kit and a detecting method. The detecting reagent kit comprises PCR mixed reaction liquid, a peptide nucleic acid probe, a taqman-MGB (minor groove binder) probe, an inner control forward and reverse primer and probe, an external control forward and reverse primer and probe and a positive reference article, wherein the inner control forward and reverse primer and probe is used for K-ras gene mutation detection, and the external control forward and reverse primer and probe is used for the K-ras gene mutation detection. The PNA (peptide nucleic acid) technology and the taqman-MGB technology are combined, relevant ARMS (amplification refractory mutation system) primers are designed, PNA can be stably combined with wild K-ras genes 12 or 13 codon sequences, only mutation specimens can be amplified, and the K-ras gene mutation can be detected. The method has the advantages that the speed is high, simplicity and convenience are realized, the specificity is good, the sensitivity is high, and the method can be used for the clinical K-ras gene mutation screening.

The invention belongs to the field of biotechnology and clinical molecular diagnosis and in particular relates to a fluorescent genotyping detection kit and a detection method for eight human K-ras gene mutations. According to the method, an MGB (minor groove binder) probe technology is combined with a mismatch ARMS (amplification refractory mutation system) technology, an MGB probe complementary with a DNA (deoxyribonucleic acid) plus strand and an ARMS downstream primer complementary with a DNA minus strand are designed at allelic gene loci, a mismatch locus is introduced in the ARMS downstream primer, the MGB probe is overlapped with the ARMS primer by not more than 5 bp, and the eight human K-ras gene mutations are subjected to genotyping detection by four reaction pipes based on multiple fluorescent PCR (polymerase chain reaction). The method has the advantages of good specificity, high sensitivity, simplicity and quickness for operation, accuracy for genotyping, simplicity for result interpretation and the like, and can be used for detecting the clinical K-ras gene mutations and helping doctors screen the crowed with effective anti-EGFR (epidermal growth factor receptor) treatment.

Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

The invention provides a fluorescent polymerase chain reaction (PCR) kit for detecting CYP2C19 genotypes, and belongs to the field of in-vitro nucleic acid testing. The fluorescent PCR kit comprises PCR reaction liquids for detecting CYP2C19*2 and CYP2C19*3 genotypes, Taq DNA polymerase, and uracil-DNA glycosylase, wherein the PCR reaction liquids for detecting the CYP2C19*2 and CYP2C19*3 genotypes respectively comprise PCR amplification primers, minor groove binder (MGB) probes and the like; and nucleotide sequences for detecting the CYP2C19*2 and CYP2C19*3 genotypes are shown as SEQ ID NO:3-4 and SEQ ID NO:5-6 respectively. The kit has high sensitivity and specificity, can monitor the reaction progress in real time, ensures short reaction time, avoids subsequent treatment, can avoid reaction product pollution to the greatest extent, and can replace the traditional protein detection or the common PCR detection to diagnose the CYP2C19 genotypes.

Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

Modified oligonucleotides are provided containing bases selected from unsubstituted and 3-substituted pyrazolo[3,4-d]pyrimidines and 5-substituted pyrimidines, and optionally have attached minor groove binders and reporter groups.

Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

Minor groove binder-oligonucleotide probes are provided along with methods for their use wherein the probes have an attached fluorophore which, in an unhybridized form exhibits very low background signal.

A probe-target reaction is made more recognizable by the provision of a mass-enhancing and / or evanescent-field-perturbing amplifier element which reacts uniquely with and binds to the probe-target pair to provide increased mass. Where the probe-target pair is hybridized dsDNA, a suitable mass-enhancing amplifier is anti-double stranded DNA mouse IgM. In examples with sufficient sequence pairs in the probe-target combination, a sequence-specific minor-groove-binding polyamide can be used that carries biotin which can be amplified by streptavidin in a suitable carrier. In a preferred embodiment, a plurality of probes are immobilized at the sites of a microarray, each probe being specific to a different target. Optics utilizing total internal reflection are described for observing perturbation of the evanescent field.

The invention relates to a detection kit and method for DNA methylation, especially to a detection kit and method for DNA methylation without CpG sites at flanking sequences. The kit and method employa Taqman probe binding to a minor groove binder (MGB) and can determine the methylation of both a CpG island (CGI) and isolated CpG sites in CpG open sea and the like without CpG at two flanking sequences. The kit and method provided by the invention are specific and sensitive, simple to operate, and does not require a control reaction or a fully-methylated standard DNA substance as a reference,thereby overcoming defects resulting from the control reaction and the fully-methylated standard DNA substance. In addition, the kit and method have higher repeatability and accuracy than the existingMethyLight technology.

The invention discloses a primer, a probe and a kit for detecting a mutation of a human JAK2 gene V617F. The primer comprises a mutation forward primer JW1-F and a mutation reverse ARMS primer JM1-R, and the probe comprises a detection probe JW1-P and a blocking probe JW1-B, wherein the mutation forward primer JW1-F is combined with a JAK2 gene conserved sequence; the mutation reverse ARMS primer JM1-R is specifically combined with a V617F (1849G>T) site mutation sequence to selectively amplify the mutation sequence; a 5minute end of the detection probe JW1-P is marked with an FAM (carboxy fluorescein) signal; a 3minute end of the detection probe is marked with an MGB (Minor Groove Binder); the detection probe JW1-P can be combined with an amplified fragment; a 5minute end of the blocking probe JW1-B is double-deoxidized and modified; a 3minute end of the blocking probe JW1-B is marked with the MGB; the blocking probe JW1-B can be specifically combined with a V617 site wild-type sequence to inhibit wild-type nonspecific amplification. The primer and the probe which are disclosed by the invention are high in specificity and good in sensitivity, and have high detectability of 1 percent. The kit prepared from the primer and the probe is accurate in detection result and easy in reading of the detection result, is simple and rapid to operate, and is wide in application range.

The invention discloses a capripoxvirus (CPV) Taqman-MGB (minor groove binder) probe multiple real-time fluorescence quantitative PCR (polymerase chain reaction) detection primer, a kit and a detection method. The multiple Taqman-MGB real-time fluorescence quantitative PCR detection method can be used for rapid detection of Lumpy Skin Disease Virus (LSDV), Goatpox Virus (GTPV) and Sheeppox Virus (SPPV). According to the method, on the basis of conserved domains of target sequences of the LSDV, the GTPV and the SPPV, a pair of primer and three Taqman-MGB probes are designed. The method only needs two step amplification method and simple reaction conditions for fast, efficient, specific and highly-sensitive detection of an objective target sequence, is simple in operation, does not need expensive equipment and reagents, has no technical requirement on operators, is low in detection cost and short in testing time, and can avoid cross contamination caused by agarose electrophoresis so as to improve the detection accuracy and reliability.

The invention discloses a kit for detecting K-ras gene mutation and a detection method with the kit. The kit comprises an amplification primer pair, a TaqMan-MGB (minor groove binder) probe and PNA (pentose nucleic acid), wherein the sequences of amplification primers are shown as SEQ ID NO:1 and SEQ ID NO:2; and the sequence of the TaqMan-MGB probe is shown as SEQ ID NO:3; and the sequence of PNA is shown as SEQ ID NO:4. The detection method comprises the following steps of: extracting DNA (deoxyribonucleic acid) of a sample to be detected; preparing K-ras wild and mutation plasmid standard products; and detecting the K-ras gene mutation, performing qualitative judgment on the K-ras gene mutation, drawing a standard curve, performing quantitative analysis on the mutation, and the like. The kit has the characteristics of quickness, simplicity, convenience, high sensitivity and high flux, and can be used for performing qualitative or quantitative detection on the K-ras gene mutation of clinical samples of tissue, urea, serum or plasma.

The invention relates to a marking method of oligonucleotide and discloses a co-modified nucleic acid segment of a locked nucleic acid (LNA) and a minor groove binding agent (MGB), namely, both the LNA and the MGB are marked on the same oligonucleotide chain. The invention enables that the capacity of identifying nucleotide sequence mutation and SNP of the oligonucleotide is enhanced by applying the combined modification of the MGB and the LNA.