63 results about "Mitochondrial RNA" patented technology

The invention discloses an indole-benzothiadiazole derivative as well as a preparation method and application thereof. The general structural formula of the indole-benzothiadiazole derivative provided by the invention is shown in the specification, wherein R is selected from one of methyl piperazinyl, diethylamino, azetidinyl and pyrrolidinyl. The derivative can be specifically combined and fluorescently labeled with mitochondrial RNA in cells, and has good selectivity, high fluorescence signal-to-noise ratio and excellent anti-interference capability in a complex biological system. Meanwhile, the indole-benzothiadiazole derivative provided by the invention is simple in synthetic route, easily available in raw materials, good in stability and convenient to store and use, can be specifically combined with RNA and generate strong fluorescence response, is suitable for real-time detection and imaging of in-vitro and in-vivo mitochondrial RNA, and is an efficient mitochondrial RNA micromolecular fluorescent probe.

The invention relates to a primer probe combination and a kit for detecting eDNA of Chinese sturgeons and application of the primer probe combination and the kit. Specific primers and TaqMan probes are designed for a D-loop region of Chinese sturgeon mitochondrial DNA, and efficient specific amplification and absolute quantification of Chinese sturgeon eDNA in an environmental sample are realized by optimizing a reaction system and amplification conditions. Compared with the prior art, the method is simple and convenient to operate, does not need to perform harmful sampling on Chinese sturgeon individuals, is suitable for resource monitoring and protection evaluation in a large-range and dynamic environment, and has high sensitivity, high accuracy and high practical value.

The application discloses a method, device and storage medium for detecting mitochondrial-derived nuclear genome sequences. The method comprises the following steps: aligning whole genome sequencing data to a mitochondrial reference genome rCRS, retaining the reads aligned to the rCRS, aligning the reads to a reference sequence containing 23 pairs of chromosomes and the rCRS, removing non-unique alignment reads; extracting potential junction reads, clustering the junction reads within a distance of 50 bp according to the alignment positions, locating the coordinates and directions of the integrated mitochondrial DNA fragments, and the nuclear genome integration position; searching for inconsistent alignment read pairs within 100 bp upstream and downstream of a read cluster, and counting the number of the inconsistent alignment read pairs as support information for the existence of non-ref NUMTs. The application utilizes the alignment information of the junction reads, detects the non-ref NUMTs through local assembly and clustering, reduces the false positive rate, can obtain accurate breakpoint and fragment information, is simple to operate, and has low detection cost.

Disclosed is a method for editing mitochondrial DNA (mtDNA) within a cell, which include introducing into the cell (a) a DNA cleaving enzyme targeted to the mtDNA sequence to be deleted; (b) a first DNA binding component targeted to a sequence adjacent to the 5′ end of a mtDNA sequence to be deleted; and (c) a second DNA binding component targeted to a sequence adjacent to the 3′ end of the mtDNA sequence to be deleted, where the DNA cleaving enzyme generates a double stranded break (DSB) within the mtDNA sequence to be deleted or generates a single strand nick on the light strand of the mtDNA sequence to be deleted, and wherein the mtDNA sequence between the target sequence for the first DNA binding component and the target sequence for the second DNA binding component is deleted.

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.

Provided are systems for strand-specific editing of DNA, including mitochondrial DNA in humans. The systems provided herein comprise a nickase and a single-stranded (ss) DNA deaminase each, or together, associated with a double-stranded DNA binding polypeptide such that DNA editing occurs in an editing region. Also provided herein are components of the systems for editing DNA taught herein, and methods of use thereof.

Compositions and methods relating to a pharmacological therapy for a human genetic disease, specifically mitochondrial DNA depletion-deletions syndromes, and more specifically, those related to mutations in the GUK1 gene. The pharmacological therapy involves the administration of deoxyguanosine (dG), a purine nucleoside phosphorylase (PNP) inhibitor, including but not limited to forodesine, or both.

The present invention relates to base editing of mitochondrial DNA. More specifically, the present invention relates to: a method for editing mitochondrial DNA bases by using a mitochondrial targeting sequence (MTS); and a base editing system used in the method. The base editing system comprises a DNA binding protein and a deaminase, or polynucleotides encoding these, and does not include an affinity tag or a polynucleotide encoding same. The present invention is useful for editing mitochondrial DNA bases of animal cells or plant cells.

The application discloses a single-cell mitochondrial whole-genome efficient and specific amplification method. The application provides a single-cell or multi-cell mitochondrial genome DNA high-fidelity amplification method, which comprises the following steps: taking total DNA obtained by lysing a test single cell as a template, using a chimeric primer composed of a 5' end non-template matching common sequence and a 3' end mitochondrial DNA (mtDNA) specific matching sequence, and a primer corresponding to the 5' end non-template matching common sequence of the chimeric primer, and using a high-fidelity DNA polymerase with strand displacement activity to simultaneously amplify several overlapping amplicons which completely cover the single-cell or multi-cell mitochondrial genome DNA. The application avoids various problems in the current single-cell mtDNA in-vitro amplification approach, and is a very reliable mtDNA whole-genome amplification method.

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.

This invention provides biomimetic nanopharmaceuticals for mitochondrial dysfunction, methods for producing the same, and applications. [Solution] The biomimetic nanodrug comprises an RGD-engineered exosome as a carrier, decalinium chloride modified on the surface of the carrier, and oxaliplatin encapsulated inside the carrier. The biomimetic nanodrug of the present invention (OXA@Exo-RD) protects cargoes that induce mitochondrial dysfunction in the blood circulation process, sequentially increasing the accumulation of mitochondria in the cargoes by targeting them. Cargo 1 (DQA) induces oxidative stress, triggering mitochondrial-mediated apoptosis and mitochondrial dysfunction, while cargo 2 (OXA) disrupts mitochondrial DNA and inhibits the initiation of DNA repair, thereby improving chemotherapy resistance. Both cargoes synergistically overcome CRC drug resistance and inhibit its migration.

This application provides a method for constructing single-cell libraries of biomaterials embedded in hydrogels, which can be used for constructing libraries of mitochondrial DNA and / or open chromatin regions and / or 3' transcriptome (RNA).

Provided herein are methods, compositions, systems, kits and uses for integrated, simultaneous spatial multiomic analyses of tissues and samples including chromatin accessibility epigenomic analysis, RNA expression transcriptomic analysis, cell cluster and cell lineage analysis, transcription factor motif analysis, extrachromosomal DNA analysis and mitochondrial DNA analysis.

The invention relates to a method for reducing mitochondrial DNA leakage, which comprises positioning nuclease in mitochondria so as to reduce the mitochondrial DNA leakage.

The invention relates to a kit for extracting and sequencing mitochondrial genome DNA of small arthropods, a sequencing method and application of the kit. The kit comprises a reagent A; a reagent B; a reagent C; the invention discloses a Tn5 transposase system for preparing a sequencing library. Aiming at the particularity of small arthropods, the trace mitochondrial DNA extracted from a single individual of a small insect by a one-step method can be used for library preparation and high-throughput sequencing, and a simple, convenient, efficient and universal technology is provided for mitochondrial whole genome sequencing analysis of numerous types of small insects. The sequencing data volume required by mitochondrial genome assembly by adopting the kit or the method disclosed by the invention can be reduced by dozens of times or even thousands of times compared with the data volume of the existing total DNA method, the difficulty that mitochondrial genome sequencing analysis cannot be carried out by utilizing a single small insect at present is overcome, the waste of rare insect samples can be reduced, and the cost is reduced. And the sequencing cost of a single sample is greatly reduced.

The present invention relates to a base editing system having the activity of correcting (A-to-G) a G11778A mutation in the mitochondrial DNA of LHON patients. Preferably, the base editing system of the present invention uses a base editor comprising a programmable DNA binding protein, a cytosine deaminase, and an adenine deaminase, or a nucleic acid encoding these proteins, wherein the cytosine deaminase is a double-stranded DNA-specific cytosine deaminase derived from Ruminococcus.

The present invention relates to a kit for extracting and sequencing mitochondrial genomic DNA of small arthropods, a sequencing method and the use thereof. The kit comprises a reagent A, a reagent B, a reagent C and a Tn5 transposase system for preparing a sequencing library. The provided kit or method is directed at the particularity of small arthropods and can achieve library preparation and high-throughput sequencing by using trace-amount mitochondrial DNA which is extracted from an individual small insect by a one-step method, thereby providing a simple and efficient universal technology for mitochondrial whole genome sequencing analysis of a wide variety of small insects. By using the provided kit or method, the volume of sequencing data required for mitochondrial genome assembly can be reduced by tens or even thousands folds than the volume of data of existing total DNA methods, thereby overcoming the problem of incapability to carry out mitochondrial genome sequencing analysis using individual small insects, and not only reducing the waste of precious insect specimens but also significantly reducing the sequencing cost for individual specimens.

This invention discloses an injectable hydrogel-MOF nanozyme composite formulation (G@ZnFe-Gel), its preparation method, and its application in pancreatic cancer immunotherapy. Specifically, the system is constructed from a Zn / Fe bimetallic MOF nanozyme loaded with glucose oxidase (GOD) and a Pluronic F127 / sodium alginate thermosensitive hydrogel. It exists as an injectable sol at 4°C and rapidly gels in situ at 37°C, achieving long-term retention and controlled release of the nanozyme. This system amplifies reactive oxygen species through a GOD-Fenton-like cascade catalysis, inducing ferroptosis in pancreatic cancer cells; the mitochondrial DNA released during ferroptosis is then absorbed into the ZnFe nanozyme. 2+ This invention synergistically activates the cGAS-STING pathway, promoting immune activation and transforming immune-cold pancreatic cancer into immune-hot tumors. In vitro and in vivo experiments have demonstrated that this system significantly inhibits pancreatic cancer growth, reshapes the immune microenvironment, and exhibits high biosafety and ease of preparation. This invention provides a novel, safe, and highly effective immunotherapy strategy for pancreatic cancer, featuring in situ delivery and multi-mechanism synergy, and holds promising clinical translational potential.

An in vitro construct useful for toxicity testing is provided, comprising: a three-dimensional (3D) scaffold comprising silk fibroin and having a crosslinked porous matrix; and stem cells adherent to the 3D scaffold. In some embodiments, the stem cells adherent to the 3D scaffold maintain stable mitochondrial DNA in long term culture. In some embodiments, the stem cells are urine stem cells.

This invention belongs to the field of biotechnology and relates to a method for tracking and assessing the metabolic distribution of platelets in organs based on mitochondrial DNA. Specifically, this invention constructs a humanized mouse model, injects exogenous platelets into the humanized mouse model, and sequences the specific SNVs of the exogenous platelet mitochondrial DNA to trace the metabolic distribution of human platelets or human drug-loaded platelets. The safety is assessed by analyzing the metabolic distribution when human platelets are used as a delivery system.

The invention belongs to the technical field of biological detection, and particularly relates to a micro-fluidic chip for detecting sepsis based on cfmtDNA and application. According to the micro-fluidic chip for sepsis detection based on cfmtDNA, on the basis of a traditional micro-fluidic chip structure, corresponding primers, probes and other structures are further designed for selected screening biomarkers, and the advantages of rapid diagnosis and application of mitochondrial DNA detection results can be achieved based on the micro-fluidic chip; and the clinical determination process of sepsis is more direct and quicker.

This invention discloses a nano-formulation HBMn-FA, its preparation method, and its application. The method includes: dissolving and dispersing a heme chloride alkaline solution in a PLGA / PLGA-PEG / DCM solution and sonicating it to obtain a mixed emulsion A; dissolving and dispersing an aqueous MnCl2 solution and an aqueous BSO solution in a PLGA / PLGA-PEG / DCM solution and sonicating them to obtain a mixed emulsion B; mixing the mixed emulsion A and emulsion B and sonicating them, then adding an aqueous serum albumin (BSA) solution and sonicating, stirring, centrifuging to collect microparticles, redissolving them in deionized water, adding polyethyleneimine and stirring, then adding DSPE-PEG-FA dropwise and stirring, centrifuging, and washing to obtain the nano-formulation HBMn-FA. This formulation can effectively exert ferroptosis, generate toxic ROS, trigger mitochondrial DNA release, and achieve endogenous signal specific activation of the STING pathway to exert anti-tumor therapeutic effects.

Provided are a one-step extraction kit, an extraction method, and a high-throughput sequencing method for vertebrate mitochondrial genomic DNA and the use thereof. The kit comprises: a reagent A comprising 3.25-4.75M guanidinium isothiocyanate, 0.65-0.85M sodium citrate, and 5-15v / v% sodium N-lauroyl sarcosinate as a cell lysis and equilibration reagent; a reagent B comprising sodium acetate; and a reagent C comprising phenol, chloroform and isoamyl alcohol at a volume ratio of (120-130) : (19-29) : 1. Reagents B and C are used for mixing with a sample of the total DNA from the lysed cells and undergoing phase separation so as to obtain the aqueous phase of the mitochondrial DNA.

Provided are methods of treating a subject having a Primary Mitochondrial Disorder (PMD) or Mitochondrial DNA Depletion Syndrome (MDDS) comprising administering a therapeutically effective amount of a compound listed in Table 1, Table 2, Table 3, Table 4, or Table 5, wherein the subject does not have a mutation in the DNA polymerase γ gene (POLG).

The invention discloses a primer combination, a kit and an identification method for parrot-shaped species identification, and belongs to the technical field of DNA identification. According to the primer combination provided by the invention, the long fragment amplification of the parrot-shaped mitochondrial genome can be realized through specific design, and a complete mitochondrial DNA sequence can be successfully obtained from trace and mixed difficult samples. The obtained full-length genome is rich in information amount, and related species with similar forms can be distinguished with high resolution. The kit provided by the invention can be adapted to a mainstream length reading sequencing platform and is simple to operate. The identification method for parrot-shaped species provided by the invention is efficient and reliable in process, and is especially good at treating old samples, trace samples or samples with complex components which are common in judicial evidence collection or field monitoring. By combining long fragment amplification and long read length sequencing, the finally output species identification result has the parrot-shaped parrot seed level or even subspecies level distinguishing precision, and the accuracy is high.

The invention belongs to the technical field of gene editing, and discloses nuclease fusion protein, a mitochondrial DNA base editing system and application. The invention provides a nuclease fusion protein. The nuclease fusion protein comprises double-mutation APOBEC3A, a Cas12 protein and a uracil glycosylase inhibitor, the double mutation APOBEC3A is subjected to W104A mutation and Y132D mutation; the Cas12 protein is GoCas12m, MmCas12m or LbCas12a which is subjected to double mutation, and the Cas12 protein is GoCas12m, MmCas12m or D832A mutation and E925A mutation are carried out on the LbCas12a subjected to double mutation. The invention provides a mitochondrial DNA base editing system comprising the nuclease fusion protein. According to the invention, the obstacle that conventional sgRNA is difficult to efficiently enter mitochondria is overcome, the editing efficiency of a CRISPR system in the mitochondria can be remarkably improved, and the existing efficiency of 0.03-0.23% is improved to a clinical available level (gt; 30%); the problem of bacterial toxicity in the construction process of the mitochondrial gene editing system is solved, the yield and stability of plasmids are improved, and large-scale preparation and application are facilitated; and the positioning efficiency of the editing component in the mitochondrial matrix is maximized. The invention provides a unified mitochondrial gene editing platform with high efficiency, high specificity and broad-spectrum applicability, and a theoretical basis is provided for precise targeting of disease-related mutation.

The invention relates to the technical field of derivatives with anti-tumor activity, in particular to a photoactivatable derivative with anti-tumor activity and a preparation method of the photoactivatable derivative. Comprise stereoisomers, tautomers, nitrogen oxides, deuterated compounds, hydrates, solvates, metabolites, prodrugs, pharmaceutically acceptable salts or co-crystals. The light-activated mitochondrial RNA polymerase inhibitor, especially the POLRMT inhibitor, is prepared for the first time, can treat diseases related to the inhibitor and various tumor diseases, has a good effect, and has relatively high cancer cell proliferation resistance activity and safety.

The invention discloses a bird and animal keratinized tissue mitochondrial DNA assembly method. The method comprises the following steps: acquiring second-generation sequencing data and cutting a low-quality sequence to generate a clean file; carrying out comparison until a mitochondrial reference genome is obtained, and screening a homologous sequence, so as to obtain unclassifid.sam; analyzing terminal base damage, counting significant damage and determining a parameter N; the SAM is converted into a mapped.fq.gz file; generating a pre-processing file based on N-substituted forward data 5'end front N basic groups and reverse data 3 'end front N basic groups; and iteratively assembling and outputting a complete mitochondrial DNA sequence by using a preset k value, thereby improving the speed and integrity.

The invention discloses a specific primer group, a probe group, an internal reference primer pair, a membrane gene chip, a detection kit, application and a detection method for two-end black knotweed, and belongs to the technical field of molecular biology detection.The two-end black knotweed mitochondrial DNA sequence is compared and analyzed, variety-specific variation sites of Cytb, D-loop and COI genes are screened, and the variety-specific variation sites of CYtb, D-loop and COI genes are obtained. Three pairs of specific primers and three biotin labeled probes are designed, a membrane gene chip detection platform is established, the primer amplification fragment length is 142-183bp, the membrane chip is suitable for detection of processed meat products, a biotin-streptavidin-alkaline phosphatase color development system is adopted by the membrane chip, the detection result is visual, and the detection efficiency is high. According to the invention, the Jinhua rhizoma anemonis can be accurately distinguished from commercial pigs such as Duroc, Changbai, Dabai and the like and other local pig varieties, the specificity reaches 100%, the sensitivity reaches a DNA template of 1ng, not less than 1% of Jinhua rhizoma anemonis components can be detected, and the whole detection process is completed within 3-4 hours.