249 results about "Mitochondrial RNA" patented technology

The specification provides programmable base editors that are capable of introducing a nucleotide change and / or which could alter or modify the nucleotide sequence at a target site in a double-stranded nucleotide sequence, such as, a chromosome, genome, or a mitochondrial DNA (mtDNA), with high specificity and efficiency. Moreover, the disclosure provides fusion proteins and compositions comprising a programmable DNA binding protein (e.g., a mitoTALE, a mitoZFP, or a CRISPR / Cas9) and evolved double-stranded DNA deaminase domains that is capable of being delivered to a cell nucleus and / or a mitochondria and carrying out precise installation of nucleotide changes in the target a double-stranded nucleotide sequence, such as, a chromosome, genome, or mtDNA. The fusion proteins and compositions are not limited for use with mtDNA, but may be used for base editing of any double-stranded target DNA.

The specification provides programmable base editors that are capable of introducing a nucleotide change and / or which could alter or modify the nucleotide sequence at a target site in mitochondrial DNA (mtDNA) with high specificity and efficiency. Moreover, the disclosure provides fusion proteins and compositions comprising a programmable DNA binding protein (e.g., a mitoTALE, a mitoZFP, or a CRISPR / Casp) and double-stranded DNA deaminase that is capable of being delivered to the mitochondria and carrying out precise installation of nucleotide changes in the mtDNA. The fusion proteins and compositions are not limited for use with mtDNA, but also may be used for base editing of any double-stranded target DNA.

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 discloses an endangered animal identification system and method based on a local comparison algorithm and a related device, and relates to the technical field of endangered animal identification, the system comprises a database which is a domestic database edited by experts, and a visual platform module receives a to-be-detected local mitochondrial DNA sequence of a to-be-detected endangered animal input by a user, the local comparison module adopts a local comparison algorithm to locally compare the to-be-detected local mitochondrial DNA sequence with a plurality of reference local mitochondrial DNA sequences in a database to obtain a local comparison result, and determines species of the to-be-detected endangered animal based on the local comparison result; and the visual platform module also displays the species of the to-be-detected endangered animal and detailed information of the species of the to-be-detected endangered animal. According to the application, the comparison efficiency and the comparison precision of the DNA sequence can be improved, and a domestic database is adopted, so that the data security and reliability are improved.

The invention provides a recombinant salmon mitochondrial gene (mDNA) as well as construction and application of a cloning vector of the recombinant salmon mitochondrial gene (mDNA). After the recombinant salmon mitochondrial gene (mDNA) is subjected to vector construction, cell transformation and proliferation amplification, polydeoxyribonucleotide (PDRN) with higher purity and yield can be prepared on a large scale, and compared with PDRN prepared from wild type mitochondrial DNA, the PDRN prepared from the recombinant mitochondrial gene has the advantages of oxidation resistance, inflammation resistance, improvement of cell proliferation ability, improvement of cell proliferation ability and the like. Compared with the prior art, the PDRN prepared from the salmon testis has remarkable and better effects in the aspects of resisting ultraviolet damage, perfecting the cell repair capacity and the like, the effect is equivalent to that of PDRN prepared from the traditional salmon testis, a new thought or method is provided for large-scale preparation or industrial production of polydeoxyribonucleotide, and the application prospect is wide.

Disclosed are a probe method-based kit and method for tracking a behavior trajectory of Acipenser dabryanus Dumeril released upstream of the Yangtze River. By analyzing mitochondrial DNA sequences of Acipenser dabryanus Dumeril and common fish species in the Yangtze River basin, a pair of specific primers ND5-F4 / ND5-R4 capable of specifically amplifying mitochondrial DNAs of Acipenser dabryanus Dumeril and a Taqman fluorescent probe ND5 Probe 4 are designed and screened. A kit composed of the specific primers, the Taqman fluorescent probe, and a ddPCR master mix is used to carry out ddPCR testing on environmental DNA samples collected before and after release, so that a behavior trajectory of the released Acipenser dabryanus Dumeril can be tracked and monitored, achieving "non-contact" tracking of the behavior trajectory of the released Acipenser dabryanus Dumeril, avoiding damage to fish bodies in the tracking process, leading to reduced tracking costs and improved accuracy of overall tracking.

Provided is a hydrogel with a biological material embedded therein, which comprises a core gel material with a biological material embedded therein, the biological material being a biological material subjected to permeabilization treatment. Enveloping biomolecules in a hydrogel and performing permeabilization treatment therein can improve the intensity of permeabilization and compatibility to various bioreactions of the biomolecules. Also provided is a method for constructing a single cell library for the biomaterial in the hydrogel with the biological material embedded therein. The method can be used for library construction for mitochondrial DNAs and / or library construction for open chromatin regions and / or library construction for 3' end transcriptome (RNAs).

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 invention provides a compound for releasing a urine fluorescent biomarker under induction of mitochondrial DNA mutation as well as a preparation method and application of the compound, and belongs to the technical field of medicines. The compound disclosed by the invention is prepared by the following steps: complementarily pairing a hybrid nucleic acid nano-carrier containing single-stranded DNA (Deoxyribose Nucleic Acid), an annular fluorescent reporter molecule and a Cas12a / crRNA compound through bases of DNA or RNA, and then coating polyethyleneimine and hyaluronic acid. A CRISPR / Cas12a system is selected as a converter of tumor specific mitochondrial DNA mutation information and urine fluorescence signals, a nano delivery system which responds to mtDNA single base mutation and is used for tumor progress monitoring and early metastasis warning is designed and synthesized, and the nano delivery system has a good application prospect in tumor progress monitoring and early metastasis sensitive detection.

An Interferon Gamma Inducible Protein 16 (IFI16) mutant gene and its use as a marker for predicting, diagnosing, or prognosticating risk or severity of chronic liver disease is described. As a result of performing genomic analysis on NAFLD and NASH patient groups, it was confirmed that the frequency of IFI16 single-nucleotide variants (SNVs) including rs2276404, rs73021847, rs7532207, and rs6940 was increased, and the expression of the IFI16 mutant gene was increased depending on the disease stage of liver disease. The IFI16 SNV was highly expressed in infiltrating macrophages, playing a role in macrophage-induced inflammatory processes, and the IFI16 variant bound more strongly to dsDNA than wild-type IFI16, exacerbating the impaired mitochondrial DNA-sensing response signaling of the IFI16-PYCARD-CASP1 pathway. Thus, the IFI16 mutant gene may be used for predicting, diagnosing, or prognosticating risk or severity of chronic liver disease.

The invention provides a kit for detecting Leber hereditary optic neuropathy and application, and relates to the technical field of biology. The kit comprises: (1) an alkaline lysis solution for releasing mitochondrial DNA in a blood sample; (2) an RPA isothermal amplification reaction system for amplifying the target sequence, wherein the RPA isothermal amplification reaction system comprises an RPA amplification primer; (3) a CRISPR-Cas12a (Clustered Regularly Interspaced Short Palindromic Repeats / Cas12a) system for carrying out high-specificity cutting on a mutation site, wherein the CRISPR-Cas12a system comprises crRNA (Complementary Ribonucleic Acid); and (4) lateral flow chromatography test paper for realizing visual detection. By simplifying the detection process, the kit greatly improves the accessibility of gene diagnosis, so that the gene screening technology can break through the limitation of traditional equipment and is popularized to a wider application scene, and the development of the gene diagnosis technology in the direction of portability, low cost and high precision is promoted. The innovation not only brings a convenient detection tool for gene mutation screening, but also lays a foundation for future gene therapy and personalized medical treatment.

The invention discloses siRNA (small interfering Ribonucleic Acid) nanoparticles capable of efficiently targeting mononuclear phagocytes and mediated by intercellular effect of neutrophils as well as a preparation method and application of the siRNA nanoparticles. The siRNA nanoparticles comprise mesoporous silica nanoparticles carrying siRNA, and functional cell membranes over-expressing ICAM-1 ligands are wrapped outside the mesoporous silica nanoparticles carrying siRNA. Key sentinel cells (mononuclear phagocytes, MPs) for targeted mitochondrial DNA release to treat inflammatory responses caused by mtDNA. The efficient delivery of MPs is realized by using a functionalized cell membrane of an overexpressed ICAM-1 ligand to wrap nano-particles and by virtue of infiltration and rapid apoptosis of a large number of neutrophils in an inflammatory microenvironment and a process (cell burial effect) of specific phagocytosis of the neutrophils by the MPs. Compared with a traditional targeting scheme, the in-vitro targeting efficiency and the in-vivo targeting efficiency of the FCM-coated siNPs scheme are improved by 360% and 120% respectively. The siRNA is guided to target MPs, and the expression of STING is inhibited, so that the inflammatory reaction caused by mtDNA release is regulated and controlled, and an excellent anti-inflammatory curative effect is shown in an inflammatory model mouse.

The invention provides a method for constructing a single cell library aiming at a biological material in hydrogel embedded with the biological material. The method can be used for carrying out library construction on mitochondrial DNA and / or carrying out library construction on a chromatin open interval and / or carrying out library construction on a 3 '-terminal transcriptome (RNA).

The present invention relates to an application and product of mitochondrial DNA G-tetramer in sperm in male infertility detection, and belongs to the field of biomedical technology. The level of mitochondrial DNA G-tetramer in sperm is used as a reference indicator for semen analysis, and is applied to male infertility detection products, providing an effective theoretical reference for male infertility in sperm pathophysiology; then, an adapted TPE-mTO fluorescent probe is used as a relevant detection means to detect the level of mitochondrial DNA G-tetramer in sperm, thereby achieving efficient and convenient clinical detection. In addition, a mitochondrial autophagy inducer is used to restore abnormal fertilization caused by increased sperm mitochondrial DNA G-tetramer levels, and the application and product of the mitochondrial autophagy inducer in abnormal fertilization are proposed, providing a new potential treatment reference and theoretical basis for patients with sperm mitochondrial DNA G-tetramer dysfunction.

Embodiments of systems, methods, and compositions provided herein relate to assays for selectively controlling enzymatic reactions. Some embodiments relate to methods of inhibiting, reducing, or eliminating secondary DNA (such as mitochondrial DNA) sequencing reads from open chromatic sequencing, whole genome sequencing, or targeted sequencing.

The invention relates to the technical field of polypeptide drugs, in particular to a nanoscale mitochondrial DNA scavenger and application thereof. The polypeptide targeting myocardial cells in the nanoscale mitochondrial DNA scavenger provided by the invention can target the myocardial cells, so that the STING inhibitor is enriched in the myocardial cells; the mtDNA probe can be combined with mtDNA in a targeted manner to form an NS-mtDNA compound; the self-assembly peptide is a convertible self-assembly unit, forms a nano structure, can transport an NS-mtDNA compound to lysosome to promote degradation, can prolong the retention time and stability of the nano-scale mitochondrial DNA scavenger and continuously release an STING inhibitor, the released STING inhibitor can inhibit an STING pathway, and the self-assembly peptide can be used for inhibiting the activity of the nano-scale mitochondrial DNA scavenger, so that the activity of the nano-scale mitochondrial DNA scavenger can be improved, and the activity of the nano-scale mitochondrial DNA scavenger is improved. Therefore, myocarditis or inflammations related to myocarditis are improved, and a new technical support is provided for treatment of myocarditis.

The present disclosure is in the field of low depth whole genome sequence and genetic sequencing. In particular, method of detecting contamination based on haplotype detection of the mitochondrial DNA.

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.

A novel genetic circuit called MitoFail (mitochondrial female-associated lethality) that supports an animal-universal female-specific repressible lethality. This genetic circuit is built around universally active restriction endonucleases shredding mitochondrial DNA molecules (mtDNA) only in females. The essential nature of mtDNA for the animal viability and yet its vulnerability to the huge diversity of mitochondria-imported restriction endonucleases support the universal adaptability of MitoFail to divergent animal species. Since MitoFail is active only in females, it is passed down and spread by competitive males and results in the suppression of a targeted population by reducing female reproductive output. MitoFail offers multiple advantages over current genetic methods of insect control and is a promising addition to the genetic toolbox for controlling diverse animal species.

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.

Provided herein are compositions and methods to assess the genomic landscape of fixed cells using light activated oligonucleotides that can be directed to the nucleus, mitochondria, or cytoplasm of fixed cells and that, upon activation, can be extended for in situ copying of nuclear single-stranded DNA (i.e., open chromatin), open mitochondrial DNA, and / or cytoplasmic RNA into barcoded complementary DNA. These methods also provide for gene specific 3D chromatin structural niche analysis.

The present disclosure provides methods for detecting macro-heteroplasmy and / or micro-heteroplasmy in mitochondrial DNA. The methods can include detecting or monitoring the presence of heteroplasmy, and / or identifying a threshold level of heteroplasmy. In addition, the methods can be used for diagnosing a mitochondrial related disease or disorder, as well as for monitoring the efficacy of a therapy affecting mitochondrial DNA (mtDNA) in a subject having or suspected of having heteroplasmy.

The invention discloses a ruthenium complex for inducing mitochondrial DNA polycondensation to overcome tumor drug resistance, an anti-tumor drug and a preparation method and application of the ruthenium complex and the anti-tumor drug. The ruthenium complex is connected with a triphenylphosphine structure-modified bipyridine diazo-anthracene ligand through an alkyl chain, so that the fat solubility of the complex is improved, and the complex can enter cells more easily; the complex has a mitochondrial enrichment function and can be enriched in mitochondria; the complex induces mitochondrial DNA polycondensation so as to cause cancer cell death; the targeting property on tumor cells is effectively improved; the IC50 of the compound on human non-small cell lung cancer cell strains and cis-platinum drug-resistant strains thereof is far lower than that of cis-platinum, and the compound can be used as an excellent anti-tumor drug. The complex takes a ruthenium-coordinated ruthenium precursor compound as a raw material, and the raw material is economical and easy to obtain; the selectivity of the ruthenium complex can be improved, and the cost is saved.

Single-cell methods are provided for investigating DNA and RNA interactions with regulatory proteins and evaluating the cytotoxicity of immune cells towards target cells, such as tumor cells. The methods utilize droplet microfluidics and imaging-based technology to facilitate the isolation and analysis of protein-bound nucleic acids from individual immune cells or apoptotic tumor cells. The methods utilize fluorescence-activated droplet sorting and are capable of isolating and characterizing mitochondrial DNA from cytotoxic immune cells.

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.

The present invention provides novel isoquinolinone compounds that are inhibitors of mitochondrial RNA polymerase for treating various diseases such as cancer and others associated with metabolic disorders and mitochondnal dysfunction.

The invention discloses a kit and a method for simultaneously capturing gene expression, chromatin openness and mitochondrial information in the same cell at the single cell level and application of the kit and the method. According to the invention, the cell membrane, the nuclear membrane and the mitochondrial membrane can be permeabilized under the condition of ensuring the cell integrity, so that various reagents for Tn5 enzyme transposon can smoothly enter the cell nucleus and the mitochondrial, and the Tn5 enzyme can cut chromatin in the cell nucleus and mitochondrial DNA in the mitochondrial and insert a linker sequence; and in the process, the RNA in the cells is ensured not to flow out and the integrity of the RNA is ensured. Based on a transposon reaction of Tn5 enzyme, a library which can be used for transcriptome and chromatin accessibility and mitochondrial information sequencing is obtained through simple PCR amplification and fragment screening, so that the method can be used for constructing a transcriptome library with an extremely low sample size, and can be combined with a single cell sorting platform based on a microfluidic or porous plate technology and the like to obtain a single cell sequence library for sequencing the transcriptome and chromatin accessibility and mitochondrial information. And further constructing the single-cell multi-omics library.

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.

A small molecule compound DHN and a preparation method and application thereof relate to the technical field of biological medicine.The compound DHN can induce the opening of a mitochondrial membrane permeability transition pore (mPTP), promote the release of mitochondrial DNA (mtDNA) into the cytoplasm, activate a cGAS-STING signal pathway, cut pyroptosis execution protein GSDME, induce pyroptosis death of melanoma cells, and provide a new target and a lead compound for the treatment of melanoma.