217 results about "Locked nucleic acid" patented technology

The present invention relates to a new method for the synthesis of purine LNA (Locked Nucleic Acid) analogues which provides a higher overall yield. The method comprising a regioselective 9-N purine glycosylation reaction followed by a one-pot nucleophilic aromatic substitution reaction of the 6-substituent in the purine ring and simultaneous nucleophile-induced intramolecular ring closure of the C-branched carbohydrate to form novel purine LNA analogues. The novel strategy is illustrated by the synthesis of the novel compound (1S,3R,4R,7S)-7-benzyloxy-1-methanesulfonylmethyl-3-(guanin-9-yl)-2,5-dioxabicyclo[2.2.1]heptane which is easily converted into (1S,3R,4R,7S)-7-hydroxy-1-hydroxymethyl-3-((2-N-isobutyrylguanin-9-yl)-2,5-dioxabicyclo[2.2.1]heptane after isobutyryl protection of the 2-amino purine group and subsequent substitution of 1-methanesulfonyl with benzoate, debenzoylation and debenzylation.

The invention relates to a novel strategy for the synthesis of Locked Nucleic Acid derivatives, such as α-L-oxy-LNA, amino-LNA, α-L-amino-LNA, thio-LNA, α-L-thio-LNA, seleno-LNA and methylene LNA, which provides scalable high yielding reactions utilizing intermediates that also can produce other LNA analogues such as oxy-LNA. Also, the compounds of the formula X are important intermediates that may be reacted with varieties of nucleophiles leading to a wide variety of LNA analogues.

The invention relates to methods for inhibiting, cloning, modifying or labelling an endogenous DNA sequence using compositions comprising recombinases in combination with exogenous polynucleotides containing “anchoring” or “locking” sequences. The anchoring sequences serve to stabilize structures formed by the exogenous polynucleotides and the endogenous DNA. The stabilized structure thus can either serve to regulate gene transcription or replication, or can allow the endogenous sequences to be labelled or pulled out, i.e. cloned, or modified.

A novel class of pharmaceuticals which comprises a Locked Nucleic Acid (LNA) which can be used in antisense therapy. These novel oligonucleotides have improved antisense properties. The novel oligonucleotides are composed of at least one LNA selected from beta-D-thio/amino-LNA or alpha-L-oxy/thio/amino-LNA. The oligonucleotides comprising LNA may also include DNA and/or RNA nucleotides.

In one aspect, the present invention provides methods for amplifying a microRNA molecule to produce DNA molecules. The methods each include the steps of: (a) using primer extension to make a DNA molecule that is complementary to a target microRNA molecule; and (b) using a universal forward primer and a reverse primer to amplify the DNA molecule to produce amplified DNA molecules. In some embodiments of the method, at least one of the forward primer and the reverse primer comprise at least one locked nucleic acid molecule.

The invention relates to the field of molecular biology and aims to provide an ARMS-qPCR (Allele Refractory Mutation System-quantitative Polymerase Chain Reaction) detection kit for KRAS (Kirsten Rat Sarcoma Viral Oncogene Homolog) gene mutation subtype and a detection method. The kit comprises a qPCR hybrid reaction solution, a locked nucleic acid retardant probe, a reference primer, an ARMS primer and a positive control sample, wherein the qPCR hybrid reaction solution comprises a PCR buffer solution, dNTPs (Deoxynucleotide Triphosphates), MgCl2, GoldStarbest Taq enzyme, a universal PCR reverse primer and a universal TaqMan probe. The kit provided by the invention can be used for rapidly and accurately detecting specific locus mutation of KRAS genes in various cancer tissues with high sensitivity, has high sensitivity, and can be used for detecting genome DNA with various tissue origins, specially free DNA segments adopting cell-free systems, such as blood serum and blood plasma, orother body fluid origins, wherein the genome DNA is derived from cell systems. Compared with direct sequencing and other mutation detection technologies, the kit and the detection method thereof havethe advantages of strong specificity, high sensitivity, simplicity and rapidness in operation, high throughput, safety, definiteness and objectivity in result identification and the like for detecting the KRAS gene mutation.

The present invention discloses a method of detecting a wild-type or mutant B-RAF gene in a body fluid sample from a subject. Also disclosed are methods of using B-RAF as a biomarker for detecting cancer, predicting the outcome of cancer, and monitoring the treatment of cancer or the status of cancer. Furthermore, the invention discloses methods and compositions for detecting a mutant gene with a peptide nucleic acid clamp capable of hybridizing to a wild-type gene and a locked nucleic acid probe capable of hybridizing to a mutant of the gene.

The invention discloses a group of probes, a detection kit and a detection method for detecting thalassemia gene point mutation based on a liquid chip of locked nucleic acid sensibilization. Due to the characteristics that locked nucleic acid can flexibly modify probes, probes for detecting thalassemia gene point mutation are designed; these probes are respectively coupled to different numbers of microspheres; a specific primer disclosed by the invention is designed to amplify mutation sites, so as to obtain a PCR product; the PCR product is hybridized with the microspheres coupled to the probes; fluorescence labeling is carried out by using a fluorescence labeling reagent; and finally detection on the mutation sites is carried out through a liquid chip detector. The probes based on LNA sensibilization are shortened in comparison with a conventional probe, and still can keep relatively high Tm values; so that the hybridization signal ratio of completely matched probes and mismatched probes is improved to over four times from below twice in the past, thus the probes are easy to distinguish.

The present disclosure provides meroduplex ribonucleic acid molecules (mdRNA) capable of decreasing or silencing ERBB gene expression. An mdRNA of this disclosure comprises at least three strands that combine to form at least two non-overlapping double-stranded regions separated by a nick or gap wherein one strand is complementary to an ERBB mRNA. In addition, the meroduplex may have at least one uridine substituted with a 5-methyluridine, a nucleoside replaced with a locked nucleic acid, or optionally other modifications, and any combination thereof. Also provided are methods of decreasing expression of an ERBB gene in a cell or in a subject to treat an ERBB-related disease.

The present invention relates to chemical modification motifs for oligonucleotides. The oligonucleotides of the present invention, such as chemically modified antisense oligonucleotides, can have increased in vivo efficacy. The chemically modified oligonucleotides provide advantages in one or more of potency, efficiency of delivery, target specificity, toxicity, and/or stability. The chemically modified oligonucleotides have a specific chemical modification motif or pattern of locked nucleic acids (LNAs). The oligonucleotide (e.g. antisense oligonucleotide) can target RNA, such as miRNA or niflNA. Also provided herein are compositions comprising the chemically modified oligonucleotides and methods of using the chemically modified oligonucleotides as therapeutics for various disorders, including cardiovascular disorders.

The invention provides an electrochemical DNA biosensor for detecting the BCR/ABL fusion gene of chronic myeloid leukemia (CML), which comprises the following steps of: (1) designing and synthesizing the specificity sequence of the CML according to the fusion locus of the universal primer sequence of the selected CML gene type to be detected; and (2) building the electrochemical DNA biosensor by combining a chemical bonding technology, a molecular hybridization technology, a locked nucleic acid probe technology with an electrochemical enzyme linked immunity technology for detecting the BCR/ABL fusion gene of the CML. The biosensor greatly strengthens the sensitivity and the specificity, thereby being capable of realizing the early diagnosis of the chronic myeloid leukemia (CML).

Provided herein are compositions and methods for next generation sequencing using universal polynucleotide adapters. Further provided are universal adapters using locked nucleic acids or bridged nucleic acids. Further provided are barcoded primers of reduced length for extension of universal adapters. Further provided herein are universal adapter blockers.

The invention discloses a primer probe group for detecting rs1045642 and application of the primer probe group. The invention firstly protects the primer probe group which comprises a primer F, a primer R and a probe P; the primer F is a single-strand DNA molecule shown as a sequence 1 in a sequence table; the primer R is a single-strand DNA molecule shown as a sequence 2 in the sequence table; the probe P is a single-strand DNA molecule with the one terminus having a fluorescent group and the other terminus having a fluorescence quenching group; a part of nucleotides in the DNA molecule are locked nucleic acids; and a nucleotide sequence of the DNA molecule is shown as a sequence 3 in the sequence table. The primer probe group provided by the invention can be used for detecting rs1045642and judging a genotype of a sample to be detected based on the site so as to guide medication and having an important application and popularization value.

The invention discloses a probe and a primer sensitized by locking nucleic acid and used for detecting C677T mutation of an MTHFR gene, a kit and a detection method. By means of the property that the locking nucleic acid can flexibly modify a TaqMan probe, a proper position of the probe used for detecting the C677T mutation of the MTHFR gene is modified by the locking nucleic acid to increase the specificity. Fluorescence quantitative PCR amplification is carried out on a sample genome DNA obtained in step (1) by the probe and the primer sensitized by the locking nucleic acid and used for detecting the C677T mutation of the MTHFR gene, in order to measure a Ct value of a wild probe and the Ct value of a mutant probe; positive is checked when the Ct value is smaller than 35 recurring numbers; only the sample detected to be positively by the wild probe is wild homozygotic type CC; only the sample detected to be positively by the mutant probe is mutant homozygotic type CC; the sample detected to be positively by both of the wild probe and the mutant probe is CT heterozygote.

The invention discloses a kit for detecting EGFR gene mutation and a detection method thereof. The kit comprises a specific probe modified with LNA locked nucleic acid. The specific probe aims at an EGFR gene mutation site. The specific probe can be combined with a wild type DNA and can detect a DNA sample contained with 0.01 percent of the EGFR gene mutation. The detecting method has the advantages that the specificity is strong, the sensitivity is high, pollution is small, the operation is simple and quick, security is high and the like. The detection method is especially suitable for detecting the gene mutation from body fluid contained with low-content mutation like blood plasma, urine and saliva and is suitable for conducting early screening and diagnosis on lung cancer, and guidance is provided for individualized medication.

Antisense oligomers useful for modulating hepatitis B virus infections, and for the treatment of hepatitis B virus (HBV) and hepatitis B virus-related conditions in animals including humans. More particularly, antisense oligomers with modified nucleotides for treatment of HBV in animals, more particularly antisense oligomers comprising 2′O-4′C-methylene-bridged sugars, or nucleotides with other 2′O-4′C bridged sugars, also known as locked nucleic acids (LNA), for treatment of HBV in animals, and more particularly for treatment of HBV in humans.

This application discloses methods for cDN′A synthesis with improved reverse transcription, template switching and preamplification to increase both yield and average length of cDNA libraries generated from individual cells. The new methods include exchanging a single nucleoside residue for a locked nucleic acid (INA) at the TSO 3′ end, using a methyl group donor, and/or a MgCb concentration higher than conventionally used. Single-cell transcriptome analyses incorporating these differences have full-length coverage, improved sensitivity and accuracy, have less bias and are more amendable to cost-effective automation. The invention also provides cDNA molecules comprising a locked nucleic acid at the 3′-end, compositions and cDNA libraries comprising these cDNA molecules, and methods for single-cell transcriptome profiling.

The invention relates to the field of gene detection, in particular to a nucleic acid membrane strip and a kit for hereditary hearing loss gene detection. The kit comprises a PCR reaction liquid, wherein the PCR reaction liquid comprises a PCR reaction liquid A, a PCR reaction liquid B, a PCR reaction liquid C, a PCR reaction liquid D and a PCR reaction liquid E and also comprises corresponding primers and probes. According to the kit, the specific amplification LATE-PCR special primers are designed by use of the sequence of various mutation sites, reported by in literature, and the locked nucleic acid modified ZNATM probes are adopted, so that the least tubes of the reaction liquid can detect the most sites. The PCR reaction liquids A, B, C, D and E in the kit can be tested on the same fluorescent PCR instrument by use of the same amplification program, and the clinical requirement for quick and convenient hearing loss detection is satisfied.