60 results about "A deoxyribonuclease" patented technology

The invention discloses a novel PDRN, a supramolecular liposome containing the PDRN as well as a preparation method and application of the supramolecular liposome. The preparation method of the novel PDRN comprises the following steps: (1) mixing a tissue raw material and PBS, crushing, carrying out enzymolysis, adding SDS, a Tween20 solution, polyethylene glycol octylphenol ether, a TE buffer solution and NaCl, stirring, centrifuging, mixing a supernatant with absolute ethyl alcohol, and standing to obtain a DNA crude extract; (2) carrying out breaking shearing treatment by using an ultrasonic breaking instrument; and (3) carrying out enzymolysis treatment by using restriction endonuclease and deoxyribonuclease, and the like. The PDRN supramolecular liposome is obtained by wrapping the PDRN with fatty acid extracted from marine microalgae. The fish testis are used as the raw material source, the selection range of the raw materials is widened, the preparation cost is reduced, and the organic reagent used in the preparation method is ethyl alcohol which is easy to remove and harmless.

The invention discloses an extraction method and application of high-molecular-weight turbellarian worm genome DNA, and belongs to the field of turbellarian worm genome extraction.The turbellarian worm genome DNA extraction is achieved by improving a lysate formula and removing ethylenediaminetetraacetic acid and increasing Mg < 2 + > on the basis of a conventional reagent in a classic Tris-HCl-phenol-chloroform method to inhibit activity of deoxyribonuclease II in a turbellarian worm body to inhibit degradation of genome DNA, and the high-molecular-weight turbellarian worm genome DNA is obtained. Meanwhile, ribonuclease A is added, RNA in cells can be degraded in the tissue cell digestion process, so that the step of two times of purification is omitted, the loss of DNA yield is reduced, experimental steps are simplified, especially the step of DNA purification is reduced, the problem of degradation in the extraction process of the turbellarian turbellarian worm genome DNA is solved, and the genome DNA yield (gt; 10 micrograms per turbellarian worm), and the turbellarian worm genome ultra-long sequencing or PacBio high-fidelity sequencing requirements are met.

The invention discloses a preparation method of a seawater shellfish single-cell suspension suitable for dynamic detection. The preparation method comprises the following steps: extracting gill filament tissues from living shellfish; washing gill filaments, and absorbing moisture; cutting the gill filament tissue into pieces; carrying out enzymolysis digestion on the gill filament tissue fragments by using an enzyme mixed solution consisting of collagenase II, hyaluronidase and deoxyribonuclease I; centrifuging and resuspending the cells; and filtering to obtain a single-cell suspension. Aiming at the condition that the shellfish tissue structure is relatively loose, cleaning liquid, enzyme liquid components, enzymolysis time, centrifugal speed and the like are optimized, and the high-quality single-cell suspension is obtained. According to the preparation method of the single-cell suspension, non-lethal sampling of seawater shellfish is conducted, the requirement for dynamic detection of shellfish traits in the industry can be met, enzymolysis is more sufficient, experiment time is shorter, operation steps are simple and convenient, obtained cells are more complete in morphology, the caking rate is extremely low, and the living cell rate is higher; and the requirements of construction of various single cell sequencing platform libraries can be fully met.

The invention relates to AAV gene therapy vectors containing novel deoxyribonuclease (DNase) transgene for delivery and expression of an enzyme which has a deoxyribonuclease (DNase) activity for enhanced clearance of cell-free DNA (cfDNA) accumulated intravasculary and / or extravasculary for treatment of various diseases and conditions, including cancer and neurodegeneration.

A method of decellularization of animal tissue, comprising the steps of: mechanical homogenization of animal-derived tissue, incubation of the homogenized tissue from step a) in a solution of trypsin and EDTA, subjecting the filtered tissue to incubation in a PBS buffer solution with a pH in the range of 7.2-7.6 containing a non-ionic detergent at a concentration of 4 - 2% and NH4OH at a concentration of 0.2 - 0.08%, rinsing the tissue with a PBS buffer solution containing an antimicrobial agent, incubation in a deoxyribonuclease solution at a concentration of 0.001 to 0.003% (w / v), rinsing the decellularized tissues obtained with a PBS buffer solution containing an antimicrobial agent, wherein the non-ionic detergent is a compound with a molecular weight of 596 g / mol and additional steps of incubating the tissue in the solution PBS buffer with a pH of 7.2- 7.6 containing a non-ionic detergent at a concentration of 1.5-0.5% and NH4OH ata concentration of 0.2-0.08%, where the incubation is carried out at a temperature below room temperature for 36-48h, and incubation of the tissue in a PBS buffer solution with a pH of 7.2-7.6 containing a non- ionic detergent at a concentration of 0.75-0.25% and NH4OH at a concentration of 0.2-0.08%, where the incubation is carried out at a temperature below room temperature for 36-48h. Bioink containing decellularized tissue of animal origin.

The present application relates to a deoxyribonuclease mutant and its application in preparing PDRN. The amino acid sequence of the deoxyribonuclease mutant is shown as SEQ ID NO. 1, and the nucleotide sequence of the encoding gene is shown as SEQ ID NO. 2. The present application also provides the application of the deoxyribonuclease mutant and / or the encoding gene of the deoxyribonuclease mutant in preparing PDRN, and the specific application method. The present application is based on the deoxyribonuclease expressed by the DNASE1 gene of bovine pancreas tissue, and the amino acids at positions 69, 132 and 215 are selected for iterative saturation mutation. Compared with the wild-type enzyme, the enzyme activity of the deoxyribonuclease mutant A69H-S132R-Q215C is increased by 2.15 times, reaching 4.1x10 6 U / mL, and the DNA extraction rate of salmon PDRN prepared by using the mutant enzyme prepared by the present application is 15.2%, the DNA content is 99.1%, and the molecular weight is 1.05 million Daltons, which is significantly improved compared with the wild-type enzyme, and the PDRN in this molecular weight range has more significant biological activity.

The application provides a kit for extracting RNA and application thereof. In the application, deoxyribonuclease (DNase) is used for DNA digestion in the process of magnetic bead extraction. The reaction environment can meet the optimal reaction environment requirement of the deoxyribonuclease (DNase), and at the same time, a hydrophobic environment is provided when the magnetic beads are adsorbed, so as to ensure the yield of nucleic acid. In addition, an emulsion of an oil is added to increase the total volume during the extraction operation, so as to facilitate the convenience of automatic extraction operation of the instrument. The extracted RNA has high yield and high purity, and the method can easily realize automatic extraction of the instrument. The requirement of extracting high-purity RNA quickly and conveniently in the process of clinical practical application is solved, the difficulty and cost of extracting high-purity RNA are greatly reduced, and the extracted RNA can be well adapted to subsequent RT-PCR and RNA-seq application.

Abstract Described herein, in one aspect, are antigen presenting cells (APCs) comprising an exogenous nucleic acid encoding one or more candidate antigens, wherein the one or more candidate antigens are expressed and presented with MHC class I or MC class II molecules; a molecular reporter of Granzyme B (GzB) activity; and c) an exogenous inhibitor of caspase-activated deoxyribonuclease (CAD)-mediated DNA degradation, a CAD knockout, or a caspase knockout (e.g., caspase 3 knockout). Described herein, in another aspect, is a system for detection of recognized antigen presentation by an antigen presenting cell to a cytotoxic lymphocyte or NK cell. Abstract 2018 / 22761 oM - cell Target ml Target cell cell cell Target Target Target SUBSTITUTE SHEET (RULE 26) cell cell cell Target Target cell cell 1 / 28 my Isolate recognized cell Library of target cells target cells and displaying different Add T cells from sample sequence antigens antigens of interest. CTLs deliver cytotoxic granules to target cells displaying cognate antigen FIG. 1 PCT / US2018 / 036663 20 26 20 53 56 07 J ul 2 02 6 2 0 2 6 2 0 5 3 5 6 0 7 J u l 2 0 2 6 2 0 1 8 / 2 2 7 6 1 o M a n d m y 1 / 2 8 m y L i b r a r y o f t a r g e t c e l l s d i s p l a y i n g d i f f e r e n tA d d T c e l l s f r o m s a m p l e of interest. CTLs deliver c y t o t o x i c g r a n u l e s t o t a r g e t c e l l s d i s p l a y i n g c o g n a t e a n t i g e n P C T / U S 2 0 1 8 / 0 3 6 6 6 3

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

The present invention provides a method for reducing the amount of DNA remaining in a biopolymer so as to increase the purity of the biopolymer when producing the biopolymer by means of a microorganism. This biopolymer production method includes: an endodeoxyribonuclease treatment step for bringing an endodeoxyribonuclease into contact with biopolymer granules obtained by culturing a microorganism capable of producing a biopolymer; and a pH adjustment step for keeping the biopolymer granules under the condition of a pH of 7.0 or higher at least at one time point after the start of said endodeoxyribonuclease treatment.

The invention discloses a recombinant vector containing a human astrovirus nucleic acid fragment, which is obtained by inserting an exogenous nucleotide sequence into a prokaryotic expression vector skeleton, and the exogenous nucleotide sequence is formed by sequentially connecting the following gene sequences: an SUMO fusion tag DNA sequence, an SAP gene sequence, an MS2 phage related DNA sequence and a human astrovirus sequence fragment. The tail end of the prepared human astrovirus pseudovirus particle contains a stem-loop structure, so that the human astrovirus pseudovirus particle is not easily degraded by nuclease, and the stability of virus nucleic acid RNA (Ribonucleic Acid) is improved; meanwhile, a human astrovirus conservative RNA gene sequence is wrapped by the phage MS2 coat protein, so that the phage MS2 coat protein has relatively high ribonuclease and deoxyribonuclease resistance, and the long-term stability of virus nucleic acid RNA is enhanced. And the stability problem of the virus RNA quality control product in the production and use processes is solved.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

The present invention discloses a recombinant Vibrio natriegens and a method for promoting the secretion and expression of exogenous proteins. The recombinant Vibrio natriegens is constructed by using the Vibrio natriegens with the preservation number of CICC 10908 as the chassis cell, knocking out the deoxyribonuclease gene on its genome, and inserting a combined expression cassette of T7 RNA polymerase and aminoglycoside adenyltransferase. The recombinant plasmid carrying the exogenous protein is transformed into the above-mentioned recombinant Vibrio natriegens, and the seed liquid is inoculated into the fermentation medium. After culturing to the mid-logarithmic growth phase, IPTG and NaCl are added, and then the culture is continued to obtain a fermentation broth containing the exogenous protein. The present invention solves the problem of secreting and expressing exogenous proteins by Vibrio natriegens at the fermentation level for the first time, with a short culture time, low cost, and certain universality, providing technical guidance for the research of Vibrio natriegens engineering bacteria in the fields of protein engineering and fermentation engineering.

The invention belongs to the field of bioinformatics, and relates to a bioinformatics method for improving the number and accuracy of predictive enhancers. According to the bioinformatics method for improving the number and accuracy of the predictive enhancer, the results of deoxyribonuclease I sensitive site enrichment sequencing (DNase-seq) and micro-nuclease sensitive site enrichment sequencing (MH-seq) are obtained by using the bioinformatics method, and more and wider chromatin open regions are obtained by combining the two sequencing, so that the accuracy of the predictive enhancer is improved. Carrying out multi-omics analysis (RNA-seq) on the basis of a large number of obtained chromatin open regions in combination with transcriptome data, and predicting more enhancers than enhancers obtained through analysis by a single method; and finally, the enhancer predicted by the two methods at the same time is obtained, and the accuracy of predicting the enhancer is improved.

A pharmaceutical composition for topical wound treatment comprising one or more nitrogenous heterocyclic compound of 5 or 6 atoms with imide group; one or more deoxyribonuclease enzyme with activity pH between 4.5 and 6.5; and one or more carboxylic acid; kits and process to obtain this pharmaceutical composition and uses for wounds treatment.

The present invention provides compositions that modulate gut microbiota and / or the activity of deoxyribonucleases (e.g., in the gastrointestinal tract) and are useful in the treatment of diseases and disorders related to the activity of deoxyribonucleases including, for example, neurodegenerative, inflammatory, and metabolic diseases and cancer.

The invention relates to the technical field of biological medicines, and particularly discloses application of deoxyribonuclease I in preparation of a medicine for treating wrapped peritoneal sclerosis. The application finds that deoxyribonuclease I can inhibit METs, can effectively reduce expression of METs in peritoneal tissues of a mouse with wrapped peritoneal sclerosis, and can also significantly improve the abdominal adhesion score, peritonitis infiltration, adhesion region collagen deposition area and peritoneal thickness of the mouse with wrapped peritoneal sclerosis; according to the present invention, the target METs is introduced to treat the wrapped peritoneal sclerosis so as to improve the pathological change related to the wrapped peritoneal sclerosis, such that the new wrapped peritoneal sclerosis prevention and treatment strategy is constructed by using the target METs to treat the wrapped peritoneal

The application discloses a kind of nanoparticles for removing lung mucus and its preparation method and application, it is related to the field of biological medicine.The nanoparticles of the application are composed of inner core and shell coated on the surface of the inner core, wherein the inner core contains citric acid and photoluminescent polymer-aliphatic polyester copolymer;The shell contains deoxyribonuclease and hydrophilic polymer.The nanoparticles provided by the application have excellent biocompatibility and multifunctional integration characteristics, which realizes effective penetration of lung mucus barrier, specific degradation of DNA network, down-regulation of pro-inflammatory factors and inhibition of mucus production in sequence through cascade response mechanism, breaking through the limitations of traditional single therapy.In addition, the in vivo experimental results show that it can significantly reduce the thickness of lung mucus and the protein concentration of lavage fluid, and effectively relieve airway obstruction, and can be used for removing lung mucus.

The invention provides a preparation method of decellularized omentum majus, and relates to the technical field of biomedical materials.The method comprises the steps that in a closed container, omentum majus subjected to freeze-drying treatment is placed in a mixed solvent to be subjected to degreasing treatment, and a first intermediate is obtained; performing vacuum pulse treatment on the closed container in the degreasing treatment process; cleaning the first intermediate to obtain a second intermediate; performing circulating oscillation treatment on the second intermediate for at least one time to obtain a third intermediate; the third intermediate is cleaned in a phosphate buffer solution and then placed in a compound enzyme solution containing pancreatin and deoxyribonuclease I to be subjected to oscillation treatment, and a fourth intermediate is obtained; the fourth intermediate is sequentially washed with deionized water and a phosphate buffer solution and then placed in a sucrose solution to be soaked, then freeze-drying and sterilization are sequentially conducted, and the decellularized omentum majus is obtained. By adopting the method disclosed by the invention, the decellularized omentum majus with high purity, low residue and complete structure can be obtained.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

The invention discloses lung glial cells as well as an in-vitro culture method and application thereof. The in-vitro culture method comprises the steps that in-vitro mouse lung tissue is subjected to mechanical separation and enzymolysis digestion, a single-cell suspension is prepared, enzymolysis digestion is conducted by adding a mixed digestive enzyme solution, and the mixed digestive enzyme solution comprises type I collagenase and deoxyribonuclease I; the single cells are placed in a DMEM complete medium for subculture, the pulmonary glial cells are obtained, and the DMEM complete medium contains the heparin-binding epidermal growth factors. According to the in-vitro culture method, the lung glial cells can be successfully obtained, and the lung glial cells are high in purity, good in cell morphology and state, normal in function and suitable for follow-up cell experiments.

The present invention provides compositions that modulate gut microbiota and / or the activity of deoxyribonucleases (e.g., in the gastrointestinal tract) and are useful in the treatment of diseases and disorders related to the activity of deoxyribonucleases including, for example, neurodegenerative, inflammatory, and metabolic diseases and cancer.

The invention relates to the field of enzymatic production of deoxyribonucleoside triphosphate, in particular to a method for enzymatic synthesis of deoxyribonucleoside triphosphate. According to the method, the salmon sperm DNA is subjected to enzymolysis through deoxyribonuclease to obtain dNMP, and the enzymolysis rate reaches 99.5% or above; and carrying out catalytic reaction on the dNMP obtained by enzymolysis by utilizing adenylate kinase, thymidylate kinase, cytidine monophosphate kinase, guanylate kinase, nucleoside diphosphate kinase and acetokinase to obtain corresponding dNTP, wherein the molar conversion rate is greater than 80%. The synthesis method is simple to operate, short in production period, easy to separate and purify and low in production cost, and has important industrial application prospects.

The present invention discloses a functionalized adhesive hydrogel and its preparation method and application, belonging to the technical field of biomedical materials. The functionalized adhesive hydrogel comprises the following raw materials: methacrylated recombinant humanized type I collagen, recombinant humanized type I collagen modified with N-(2-aminoethyl)-4-(4-(hydroxymethyl)-2-methoxy-5-nitrophenoxy) butyramide, a photoinitiator and water; per milliliter of the functionalized adhesive hydrogel also comprises 80-120 U of deoxyribonuclease I, 400-600 ng of clemastine free drug and 5-15 μg of clemastine liposome. The present invention also discloses the preparation method and application of the functionalized adhesive hydrogel. The functionalized adhesive hydrogel of the present invention can reduce the occurrence of secondary injury, inflammatory response and myelin injury at the intracerebral hemorrhage site, inhibit the formation of neutrophil extracellular traps, and can achieve multi-level intervention after intracerebral hemorrhage.

The present invention discloses a novel PDRN, a supramolecular liposome containing the PDRN, and a preparation method and use thereof. The preparation method of the novel PDRN comprises: (1) mixing tissue raw materials with PBS, crushing, enzymolysis, adding SDS, Tween20 solution, polyethylene glycol octylphenyl ether, TE buffer, NaCl, stirring, centrifuging, mixing the supernatant with anhydrous ethanol, and standing to obtain a crude DNA extract; (2) using an ultrasonic interrupter to interrupt and shear the tissue; (3) using restriction endonucleases and deoxyribonucleases to perform enzymolysis, etc. The PDRN supramolecular liposome is obtained by encapsulating the above-mentioned PDRN with fatty acids extracted from marine microalgae. The present invention uses fish testis as a raw material source, broadens the range of raw material selection, reduces preparation costs, and the organic reagent used in the preparation method of the present invention is ethanol, which is easy to remove and relatively harmless.

This invention belongs to the medical field and provides a calcium carbonate manganese nanoprobe for in situ imaging detection of vascular epidermal growth factor, its preparation method, and its application. In this strategy, a DNA strand (P1 and P2) containing a protein recognition antibody, a protein recognition sequence, and a deoxyribonuclease, along with a molecular beacon DNA strand (H1), is stirred in a DMEM cell culture medium containing calcium chloride and manganese chloride to prepare the calcium carbonate manganese nanoprobe. In acidic tumor cells, this nanoprobe releases P1, P2, H1, and manganese ions. P1 and P2 can simultaneously recognize proteins, forming a complete deoxyribonuclease center, which catalyzes the cleavage of H1 under the action of the cofactor manganese ions, generating a fluorescent signal. This strategy achieves specific in situ dynamic imaging of low-abundance proteins in vivo and has great potential in biomedical research.

Therapeutic compositions for treating a subject, the compositions including a polymeric nanoparticle, an active agent encapsulated within the polymeric nanoparticle, and a delivery vehicle for targeted delivery of the polymeric nanoparticle and encapsulated agent to a target tissue in the subject. The therapeutic compositions can be designed for targeting the destruction of a Nuclei neutrophil extracellular trap (NET) in a venous thrombus in a subject. The active agent can be an enzyme, including deoxyribonuclease 1 (DNase 1) enzyme. Methods of treating a subject using the therapeutic compositions are provided, including methods of treating venous thrombus, deep vein thrombosis, or a related blood clotting disease and / or condition.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

This invention provides a Csm6-based deoxyribonuclease sensor, characterized by comprising the following components: (1) a deoxyribonuclease enzyme chain and a deoxyribonuclease substrate chain, wherein the nucleotide sequence of the deoxyribonuclease substrate chain is shown in SEQ ID No. 1 or SEQ ID No. 2 of the sequence listing, and the nucleotide sequence of the deoxyribonuclease enzyme chain is shown in SEQ ID No. 10 of the sequence listing; (2) a Csm6 protease; and (3) a single-stranded RNA reporter probe modified with fluorescent and quenching groups, wherein the sequence of the single-stranded RNA reporter probe modified with fluorescent and quenching groups is shown in SEQ ID No. 16 of the sequence listing. This invention also provides an application of this deoxyribonuclease sensor for lead detection. The deoxyribonuclease sensor provided by this invention can improve the sensitivity and selectivity of existing biosensors in lead ion detection.

The invention discloses a ternary co-modified engineered cornu cervi pantotrichum MSCs exosome and a preparation method and application thereof, and belongs to the technical field of biological medicine, the ternary co-modified engineered cornu cervi pantotrichum MSCs exosome comprises a cornu cervi pantotrichum MSCs exosome, a poly-arginine polypeptide-deoxyribonuclease I compound located on the surface of the cornu cervi pantotrichum MSCs exosome, and an RGD polypeptide modified on the surface of the cornu cervi pantotrichum MSCs exosome and containing an Arg-Gly-Asp sequence. In order to solve the problem that an integrated treatment mode of multiple pathological links cannot be realized during existing RA treatment, three active components, namely poly-arginine polypeptide (P-Arg), deoxyribonuclease I (Dnase I) and RGD polypeptide, are cooperatively modified on the surface of an exosome of pilose antler MSCs for the first time to form a surface engineering modified exosome complex; multiple functions of immunoregulation, cfDNA removal, targeted delivery and tissue protection can be realized in RA focuses, and a brand-new and systematic treatment means is provided for rheumatoid arthritis.