11 results about "Dna mutation" patented technology

Described herein is a base editing system for correcting mutations G3460A, G11778A, or T14484C in mitochondrial DNA of a patient with Leber hereditary optic neuropathy (LHON) to a normal genotype. Also, described herein is a method for correcting a mutation in the mitochondrial genes of a patient with LHON to a normal genotype using a base editor that recognizes specific sites in the mitochondrial genes of the patient with LHON and has an activity of specifically correcting the adenine base at position 3460 or 11778, or the cytosine base at position 14484, by using a fusion protein or a polynucleotide encoding such a fusion protein. The base editor or nucleotide described herein may correct DNA mutations specific to LHON in a cellular or extracellular in vitro environment. Thus, described herein is also the use of the substance in the prevention or treatment of LHON.

The present invention relates to a base correction system that corrects mitochondrial DNA mutations G3460A, G11778A, or T14484C, which are present in patients with Leber's hereditary optic neuropathy (LHON), to a normal genotype. Specifically, the present invention provides a base editor capable of correcting a mutation site in a mitochondrial gene of an LHON patient to a normal genotype. The present invention also provides a method for correcting a mitochondrial gene mutation using a fusion protein or a polynucleotide encoding such a fusion protein that recognizes a specific site in the mitochondrial gene of an LHON patient and specifically corrects the adenine base at position 3460, the adenine base at position 11778, or the cytosine base at position 14484. The base editor or polynucleotide according to the present invention can be used in cells or in an extracellular test tube environment to correct DNA mutations specifically expressed in LHON, and more preferably, can be used as a gene therapy agent to prevent or treat the disease. Thus, the present invention also provides a use of the substance for preventing or treating Leber's hereditary optic neuropathy.

The invention relates to a detection strategy of a programmable TMSD-CRISPR system for DNA / RNA (deoxyribonucleic acid / ribonucleic acid) single base mutation. Based on split crRNA and dynamic and thermodynamic regulation and control mechanisms of a dynamic nanotechnology, the purpose of identifying DNA / RNA single-base mutation at different sites is achieved by utilizing different kinetic rates of toehold-mediated strand displacement on strand displacement with a single-base mismatched target strand. According to the present invention, the identification factors for the ssDNA / dsDNA / RNA mutation are significantly enhanced (the ssDNA mutation is 169.4, the dsDNA mutation is 243.8, and the RNA mutation is 156.2), such that the DNA mutation detection with the abundance as low as 0.025% is achieved, and the single base mutation detection is allowed to be performed in the miR-21; in the research of a queue containing 75 clinical samples, the method successfully detects and accurately identifies all KRAS G12V variants at the accuracy rate of 100%, and shows the huge potential of the KRAS G12V variants in clinical molecular diagnosis. The programmable split TMSD-CRISPR has excellent specificity and adjustability, and can be used as a multifunctional platform for biomedical research, molecular diagnosis and precision medicine.

The invention discloses a lung cancer detection primer probe composition and application thereof in a diagnostic kit, and belongs to the technical field of biological detection. The technical problem to be solved is to provide a high-sensitivity and high-specificity primer probe composition for one-time detection of multiple lung cancer mutant genes. The invention provides a primer probe composition which is high in sensitivity (capable of detecting DNA mutation with mutation frequency as low as 0.5% and RNA fusion of 25 copies) and high in specificity and is used for detecting multiple lung cancer mutation genes at one time, and the primer probe composition is assembled into a kit, comprises EGFR, ALK, ROS1 and MET genes, covers 26 gene mutations and 26 gene fusion, and is used for detecting multiple lung cancer mutation genes. The detection result is high in sensitivity, strong in specificity, good in repeatability and simple, convenient and rapid to operate, and a rapid and reliable detection method is provided for clinic.

This invention relates to a system, kit, and application for the one-pot detection of miRNA and DNA mutations based on CRISPR-Cas12a and aptamers, belonging to the field of biomedical technology. The system for the one-pot detection of miRNA and DNA mutations based on CRISPR-Cas12a and aptamers includes: Cas12a protein, crRNA-R targeting miRNA, aptamer P1, aptamer P2, a linker, AO, and crRNA-D targeting DNA mutations. This invention combines CRISPR / Cas12a with a aptamer-based detection strategy to develop a label-free, direct detection system compatible with smartphones. This system can simultaneously detect EGFR mutations and miRNA-21 expression levels.

The present application relates to a primer composition, kit and method for detecting benign and malignant thyroid nodule related gene mutations, wherein the primer composition comprises primer pairs for detecting DNA mutations and RNA fusion variations of benign and malignant thyroid nodule related genes. The primer pairs can be amplified individually, and more preferably, the DNA and cDNA templates to be tested can be multiplexed with high efficiency, high sensitivity and high specificity in the same PCR reaction system and reaction program. Subsequently, by constructing a library and sequencing, the gene mutation of the relevant region can be obtained, thereby assisting the clinical diagnosis of benign and malignant thyroid nodules. Furthermore, by connecting specific nucleotide sequences to the 5' end of the upstream primer and the 3' end of the corresponding downstream primer in the primer composition in a proper proportion, the uniformity and specificity of the multiplex PCR amplification can be further improved, and the nucleic acid input for detection and the minimum detection limit of mutation frequency (as low as 0.5%) can be effectively reduced.

The application discloses a probe and a method for detecting micro residual lesions. The probe for detecting micro residual lesions is a four-layer encrypted imbricated hybrid capture probe, wherein two layers of probes are wild-type probes, and two layers are mutant probes. The nucleic acid sequence of the wild-type probe comprises the sequence described in SEQ ID NO. 1-SEQ ID NO. 74, and the nucleic acid sequence of the mutant probe comprises the sequence described in SEQ ID NO. 75-SEQ ID NO. 149. The four-layer encrypted imbricated hybrid capture probe for detecting micro residual lesions is designed, a micro residual lesion molecular detection method with high sensitivity and good specificity is provided, the low-frequency mutation capture efficiency is high, and the detection limit of the ct DNA mutation frequency containing micro residual lesions can be as low as 0.005%.

Disclosed are compositions and methods related to detecting rare mutations (e.g., somatic mutations) or genome structure variants using rolling circle-based linear amplification and next generation sequencing.

The invention relates to a system for simultaneously detecting histone modification information, DNA methylation information and DNA mutation information in circulating free DNA, belongs to the technical field of biological detection, and solves at least one of the problems that a method in the prior art cannot simultaneously detect the three types of information, and a preparation method is tedious, high in cost, large in error and the like. By optimizing the technical scheme, synchronous detection of cfDNA multi-omics information is realized, and the operation process is simplified. In the whole process, the detection time is remarkably shortened, and the detection efficiency is improved. Meanwhile, the operation steps are reduced, the possibility of sample pollution, loss and error accumulation is also reduced, and the accuracy and the stability of a detection result are ensured.

The application discloses a library construction method for simultaneously detecting various gene mutation types of brain tumors, and a composition and application thereof. The sequencing library construction method established by the application is a one-step library construction method, which places DNA mutation and RNA fusion in one PCR reaction system, and optimizes pyrophosphate detection MGMT and FISH detection CNV into an amplicon one-step method, so that DNA level mutation and RNA level fusion can be simultaneously detected. Moreover, after the library construction, QPCR quantification is not needed, and only Qubit quantification is needed, and then direct machine sequencing operation is simple. The application can be applied to rapid, simple and accurate detection of brain tumor basic items.

The present invention relates to a method for testing gene editing in an extracellular test tube environment (cell-free, in vitro system), and more specifically, to a method for confirming and quantifying target DNA mutations caused by base editors without the use of eukaryotic cells or other complicated testing processes. By using the present invention, target DNA mutations caused by base editors can be confirmed and quantified without the use of cells or other complicated testing processes. Furthermore, base editors that have the ability to correct target DNA mutations in actual cells can be screened through a simple extracellular testing method.