132 results about "Adenosine deaminase" patented technology

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. The disclosure provides fusion proteins of nucleic acid programmable DNA binding proteins (napDNAbp), e.g., Cas9 or variants thereof, and nucleic acid editing proteins such as cytidine deaminase domains (e.g., novel cytidine deaminases generated by ancestral sequence reconstruction), and adenosine deaminases that deaminate adenine in DNA. Aspects of the disclosure relate to fusion proteins (e.g., base editors) that have improved expression and / or localize efficiently to the nucleus. In some embodiments, base editors are codon optimized for expression in mammalian cells. In some embodiments, base editors include multiple nuclear localization sequences (e.g., bipartite NLSs), e.g., at least two NLSs. In some embodiments, methods for targeted nucleic acid editing are provided.

The invention relates to the field of preparation and application of biological peptides, in particular to a swelling hippocampal peptide and application thereof in preparation of uric acid reducing and gout resisting products. The amino acid sequence is shown as SEQ ID NO.1. The uric acid reducing effect of the compound is proved through in-vitro xanthine oxidase (XOD) and adenosine deaminase (ADA) inhibition experiments and cell experiments; on the basis, the uric acid reducing and gout resisting activity of the peptide sequence is further verified by means of a hyperuricemia animal model. The achievement provides a solid theoretical support for high-value development and utilization of the hippocampus japonicus and research and development of uric acid reducing and gout resisting related products.

A fusion protein comprising: a first component comprising an antibody, or a fragment or variant thereof; and a second component comprising a cytokine trap or an adenosine deaminase or a fragment or variant thereof. In certain embodiments, the antibody is an anti-PD-1 antibody. In certain embodiments, the antibody binds to a tumor antigen, for example a MUC16 or MUC1 antigen. In certain embodiments, the cytokine trap is a TGF-β trap. A polynucleotide encoding such a fusion protein and a vector comprising such a polynucleotide. A composition comprising the fusion protein. A method of using the composition, including in the treatment of cancer.

The present disclosure generally relates to an evolved cytidine deaminase derived from a cytidine deaminase, and a method of editing DNA using the same. In some aspects, the present disclosure describes directed evolution of a TadA-derived adenosine deaminase (TadA-CD) for cytidine deamination. In some embodiments, the TadA-CD comprises a plurality of mutations as compared to the parent TadA variant. In some embodiments, the TadA-CD is fused to a programmable DNA binding protein. Other aspects of the present disclosure generally relate to a cytosine base editor (CBE) comprising a programmable DNA binding protein and a TadA-CD. In some embodiments, the disclosed cytosine base editor has an improved conversion efficiency and a reduced off-target editing frequency compared to a naturally occurring CBE. Polynucleotides, vectors, and kits for generating and delivering CBE are also provided. Also provided are cells containing such vectors and CBE. In addition, methods of treatment comprising administration of CBE are also provided.

Disclosed are an aqueous American ginseng extract, a method for preparing, and use thereof in reducing uric acid. The method comprises the following steps: grinding American ginseng, mixing with petroleum ether, and subjecting the mixture to ultrasonic extraction and filtration at reduced pressure; drying the defatted American ginseng powder and mixing with deionized water, and subjecting the mixture to another ultrasonic extraction and filtration at reduced pressure to give an aqueous American ginseng extract solution; and concentrating and lyophilizing the aqueous American ginseng extract solution to give the aqueous American ginseng extract. The resultant aqueous American ginseng extract can effectively reduce the activity of xanthine oxidase and adenosine deaminase, lower the blood uric acid level, and provide certain protective effects on the kidneys, thus possessing good development prospects.

The present disclosure provides adenosine deaminase, fusion proteins, base editor systems and uses, and specifically provides an adenosine deaminase polypeptide having an amino acid sequence having at least about 70%, 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% sequence identity to SEQ ID NO: 13. Also provided are novel nucleic acid modified enzymes having improved efficiency, compositions of base editors comprising the same, and methods and uses for editing target sequences using these adenosine deaminase variants.

RNA editing tools for use in systems designed to measure RNA in vivo and manipulate specific cell types are disclosed herein. An RNA sensor system comprising a) a single-stranded RNA (ssRNA) sensor comprising a stop codon and a payload; optionally wherein the ssRNA sensor further comprises a normalizing gene; and b) an adenosine deaminase acting on RNA (ADAR) deaminase; wherein the sensor is capable of binding to a ssRNA target to form a double-stranded RNA (dsRNA) duplex that becomes a substrate for the ADAR deaminase; wherein the substrate comprises a mispairing within the stop codon; and wherein the mispairing is editable by the ADAR deaminase, which editing can effectively remove the stop codon so as to enable translation and expression of the payload. A method of quantifying ribonucleic acid (RNA) levels using the RNA sensor system is also disclosed.

The application discloses application of total flavones of chrysanthemum morifolium ramat and metabolites thereof in preparation of uric acid-lowering or liver and kidney protecting drugs or health care products. The total flavones of chrysanthemum morifolium ramat contain various flavone compounds. In a hyperuricemia model mouse, the total flavones of chrysanthemum morifolium ramat can significantly reduce blood uric acid level, and has a significant protective effect on liver and kidney, and the effect is achieved by inhibiting the activity and expression of xanthine oxidase and adenosine deaminase in the liver, and regulating the expression of uric acid related transporters in the kidney. Meanwhile, metabolites of the total flavones of chrysanthemum morifolium ramat, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide, can also play a pharmacological role in the mouse body. In vitro experiments and mouse primary hepatocytes further prove that luteolin-7-O-glucuronide and apigenin-7-O-glucuronide have an inhibiting effect on the activity and expression level of xanthine oxidase. The application widens the application range of traditional Chinese medicinal materials, and provides a new direction for development of efficient and low-toxic uric acid-lowering and anti-gout drugs or health care products.

The invention discloses application of adenosine deaminase or a modifier thereof in preparation of a medicine for treating diabetic nephropathy. After intraperitoneal injection treatment of polyethylene glycol modified escherichia coli expressed mouse adenosine deaminase is carried out on a mouse with diabetic nephropathy, the damage degree of the kidney of the mouse is obviously weakened. According to the invention, the adenosine deaminase or the modifier thereof has an obvious treatment effect on the type 2 diabetic nephropathy for the first time, and the adenosine deaminase and the modifier thereof have potential application prospects in preparation of the medicines for treating the type 2 diabetic nephropathy.

The invention belongs to the field of nucleic acid editing, and particularly relates to the technical field of regularly clustered interval short palindromic repeat (CRISPR). Specifically, the invention provides adenosine deaminase which is fused with DNA binding protein, can be used for single base editing of target nucleic acid, and has a wide application prospect.

The invention provides a composition for reducing uric acid and relieving gout and a preparation method thereof, and relates to the technical field of products for reducing uric acid and relieving gout. The composition comprises the following raw materials: 18-24 parts of phellinus igniarius, 9-13 parts of sunflower disc small molecule peptide extract, 3-7 parts of corn stigma, 4-6 parts of poria cocos, 2-5 parts of pawpaw, 3-6 parts of liquorice, 2-5 parts of astragalus extract and 3-6 parts of lettuce extract. The preparation method of the sunflower disc small-molecule peptide extract comprises the following steps: carrying out enzymolysis on sunflower disc powder twice, carrying out membrane filtration to obtain a sunflower disc small-molecule peptide solution, reacting the sunflower disc small-molecule peptide solution with chitosan oligosaccharide and transglutaminase, and carrying out freeze drying to obtain the sunflower disc small-molecule peptide extract. Through the proper raw material composition and proportion, the metabolic function can be effectively regulated to promote excretion of uric acid, so that the content of xanthine oxidase and adenosine deaminase in the liver is obviously reduced, the uric acid level is effectively reduced, the intestinal probiotic structure can be stabilized, the immunity is enhanced, the kidney is protected, and the good effects of reducing uric acid and relieving gout are achieved.

The invention relates to chemically modified oligonucleotides for use in site-directed A-to-I editing of a target RNA inside a cell with endogenous adenosine deaminase acting on RNA (ADAR), the oligonucleotide comprising a sequence capable of binding to a target sequence in a target RNA and a central base triplet (CBT) of (3) nucleotides (N-1 N0 N+1), wherein N0 is the central nucleotide directly opposite to a target adenosine in the target RNA that is to be edited, wherein the oligonucleotide comprises at least one internucleoside linkage that is a methanesulfonyl (mesyl) linkage.

The invention provides for systems, methods, and compositions for targeting and editing nucleic acids. In particular, the invention provides non-naturally occurring or engineered RNA-targeting systems comprising a RNA-targeting Cas13 protein, at least one guide molecule, and at least one adenosine deaminase protein or catalytic domain thereof.

The utility model provides a transport frame for adenosine deaminase assay kits, which comprises a trolley body, vertical plates are symmetrically arranged at the upper part of the trolley body, a plurality of groups of placing components are uniformly arranged between the two vertical plates along the vertical direction, and each group of placing components comprises a mounting plate which is connected to the opposite surfaces of the two vertical plates through a mounting part; the installation parts are used for adjusting the inclination angles of the installation plates and all parts on the installation plates, and a containing plate is arranged on the lower portions of the two installation plates. The two vertical plates are matched with the plurality of groups of placing assemblies, so that the biochemical components of the adenosine deaminase determination kit and the biochemical components in the determined reagent tube can be separately placed and stored, and the influence on the detection sensitivity and accuracy is avoided.

The present application relates to the preparation and application field of biological peptide, and more particularly to a hippocampus abdominalis peptide and its application in preparation of products for reducing uric acid and resisting gout. The amino acid sequence is shown as SEQ ID NO. 1, and the present application confirms the uric acid reducing effect through in vitro xanthine oxidase (XOD) and adenosine deaminase (ADA) inhibition experiments and cell experiments; on this basis, the uric acid reducing and gout resisting activity of the peptide sequence is further verified by means of a high uric acid animal model. This achievement provides a solid theoretical support for high-value development and utilization of hippocampus abdominalis, and research and development of products for reducing uric acid and resisting gout.

The present application provides an engineered circular RNA capable of recruiting adenosine deaminase acting on RNA (ADAR; arRNA) and its application in treating Duchenne muscular dystrophy (DMD).

The application provides an adenosine deaminase content detection method and application thereof. The adenosine deaminase content detection method comprises the following steps: using the intrinsic fluorescence of adenosine, using adenosine deaminase to hydrolyze adenosine, and detecting the content of adenosine deaminase by a fluorescence analysis method. Adenosine itself has strong fluorescence, and after being hydrolyzed by adenosine deaminase, the fluorescence of the obtained hydrolysis product is reduced, thereby causing the fluorescence intensity of the solution of the detection system to be reduced; the content of adenosine deaminase in the system is detected by the change of the fluorescence intensity before and after adenosine hydrolysis. The adenosine deaminase content detection method in the application is simple to operate, does not need large instruments and equipment, and is cheap and easy to obtain materials, can accurately and rapidly detect the content of adenosine deaminase in a sample to be detected, has good sensitivity and specificity, and has a relatively wide application prospect.

The invention discloses an adenosine deaminase mutant. The adenosine deaminase mutant is a C141R mutant and an L144P mutant based on a wild type. The invention also discloses a pichia pastoris base editor containing the adenosine deaminase mutant, the pichia pastoris base editor can realize accurate editing of specific base sites of a pichia pastoris genome, and a gene editing tool is provided for basic and application research based on pichia pastoris.

The present invention discloses a preparation method and application of a composite plant extract for inhibiting the activity of adenosine deaminase, and belongs to the technical field of plant extract preparation. The present invention relates to a process of mixing clove buds and wild chrysanthemum inflorescences, and increasing the release rate of alcohol-soluble active substances (esters and ketones) in plant cells by common steam explosion, and further increasing the purity of alcohol-soluble active substances in the composite plant extract after alcohol extraction and water washing and impurity removal steps. The composite plant extract has a good inhibitory effect on the activity of adenosine deaminase, and can effectively reduce the ammonia odor of feces of pet dogs after feeding them, and can also improve the quality of dog hair.

The invention discloses a walnut-derived xanthine oxidase inhibitory peptide and application thereof, and belongs to the technical field of bioactive peptide biology. The amino acid sequence of the xanthine oxidase inhibitory peptide provided by the invention is as shown in SEQ ID No. 1. Through in-vitro activity determination, the IC50 value of the inhibitory peptide is 3.34 mg / mL, and it is proved that the inhibitory peptide has high inhibitory activity on xanthine oxidase. Besides, the inhibitory peptide can reduce the levels of creatinine, urea nitrogen, adenosine deaminase, glutamic oxalacetic transaminase and glutamic-pyruvic transaminase, regulate the expression of kidney uric acid resorption protein URAT1, uric acid excretion protein GLUT9 and ABCG2, and significantly reduce the generation of uric acid, can be used for inhibiting xanthine oxidase and preventing or improving hyperuricemia, is applied to preparation of drugs for improving hyperuricemia, and has broad application prospects. The medicine takes the active peptide as the only active ingredient or one of the active ingredients, and has a good application prospect.

The present invention discloses an adenosine deaminase mutant, which is a C141R and L144P mutant based on the wild type. The present invention also discloses a Pichia pastoris base editor containing the adenosine deaminase mutant. The Pichia pastoris base editor can achieve precise editing of specific base sites in the Pichia pastoris genome, providing a gene editing tool for basic and applied research based on Pichia pastoris.

A fusion protein comprising: a first component comprising an antibody, or a fragment or variant thereof; and a second component comprising a cytokine trap or an adenosine deaminase or a fragment or variant thereof. In certain embodiments, the antibody is an anti-PD-1 antibody. In certain embodiments, the antibody binds to a tumor antigen, for example a MUC16 or MUC1 antigen. In certain embodiments, the cytokine trap is a TGF-β trap. A polynucleotide encoding such a fusion protein and a vector comprising such a polynucleotide. A composition comprising the fusion protein. A method of using the composition, including in the treatment of cancer.

The invention belongs to the technical field of functional microorganisms, discloses application of pediococcus pentosaceus A21243 in reducing blood uric acid and improving renal functions, and provides application of the pediococcus pentosaceus A21243 in preparing products for preventing or treating nephropathy and hyperuricemia, the pediococcus pentosaceus is the pediococcus pentosaceus A21243, and the preservation number is GDMCC NO: 63749. The Pediococcus pentosaceus A21243 has the effects of improving and delaying renal insufficiency caused by hyperuricemia, hyperglycemia and cis-platinum, and meanwhile, the Pediococcus pentosaceus A21243 can also promote excretion of uric acid through intestinal feces by inhibiting the activity of purine metabolism related enzymes in a uric acid generation pathway, such as the activity of xanthine oxidase (XOD), purine nucleoside phosphorylase (PNP) and adenosine deaminase (ADA), so that the Pediococcus pentosaceus A21243 can be used for improving and delaying renal insufficiency caused by hyperuricemia, hyperglycemia and cis-platinum. The oxidative stress disorder caused by high uric acid is regulated, so that the beneficial effect of reducing blood uric acid is achieved.

A base editing system for achieving A-to-C and / or A-to-T base mutations and a use thereof are provided. A base editor is constructed by means of fusing 3-methyladenine DNA glycosylase with adenosine deaminase and Cas9 nuclease with impaired catalytic activity, which achieves adenine-based transversion for the first time. It is found through experimental comparison that AXBE, which is constructed by means of fusing mouse-derived 3-methyladenine DNA glycosylase with adenosine deaminase TadA-8e derived from E. coli and Cas9 nickase with impaired catalytic activity derived from Streptococcus pyogenes, has the best effect of catalyzing the transversion of adenine. The use of the base editing system in the gene therapy, cell therapy, human disease model production, and crop genetic breeding, etc. is promoted.

This application belongs to the field of biotechnology and relates to an efficient and specific editing system, method, and application for A-G base substitution. The fusion protein described in this application comprises, from the N-terminus to the C-terminus, a first SaCas9 nickase fragment, a chimeric deaminase fragment, and a second SaCas9 nickase fragment. The deaminase is selected from adenosine deaminase or a variant thereof, specifically ecTadA8e. The amino acid sequence of ecTadA8e is shown in SEQ ID NO. 20, or has more than 80% sequence identity with the amino acid sequence shown in SEQ ID NO. 20, and possesses the function or activity of ecTadA8e. The fusion protein provided in this application, combined with the corresponding guide RNA, can efficiently and specifically replace the base A with G at the target site, providing an effective tool for repairing pathogenic mutations, studying gene function, and improving cell function, and has promising application prospects.

The present invention relates to methods and compositions for disrupting interaction of an NRF2 protein and a KEAP1 protein. The methods include contacting at least one polynucleotide selected from the group consisting of a polynucleotide encoding the NRF2 protein and a polynucleotide encoding the KEAP1 protein with a guide oligonucleotide that effects one or more (e.g., at least two) adenosine deaminase acting on RNA (ADAR)-mediated adenosine to inosine alterations in said at least one polynucleotide, wherein the adenosine to inosine alterations generate a mutant amino acid, thereby disrupting interaction of the NRF2 protein and the KEAP1 protein. The invention also relates to methods of treating a KEAP1-NRF2 pathway related disease in a subject in need thereof, the method comprising contacting, within the subject, at least one polynucleotide selected from the group consisting of a polynucleotide encoding an NRF2 protein and a polynucleotide encoding a KEAP1 protein with a guide oligonucleotide that effects an adenosine deaminase acting on RNA (ADAR)-mediated adenosine to inosine alteration in said at least one polynucleotide, wherein the adenosine to inosine alteration generates a mutant amino acid, thereby disrupting interaction of the NRF2 protein and the KEAP1 protein and treating the disease in the subject; and compositions thereof.

The invention relates to chemically modified oligonucleotides for use in site-directed A-to-I editing of a target RNA inside a cell with endogenous adenosine deaminase acting on RNA (ADAR), the oligonucleotide comprising a sequence capable of binding to a target sequence in a target RNA and a central base triplet (CBT) of 3 nucleotides (N−1N0N+1), wherein N0 is the central nucleotide directly opposite to a target adenosine in the target RNA that is to be edited, wherein the oligonucleotide comprises at least one internucleoside linkage that is a methanesulfonyl (mesyl) linkage.

The invention provides a double-base pichia pastoris base editor aiming at pichia pastoris, the double-base pichia pastoris base editor is a DNA molecular composition, the DNA molecular composition comprises a first expression plasmid containing a coding gene of nCas9 protein, a coding gene of pyrimidine deaminase and a coding gene of adenosine deaminase, and a second expression plasmid containing a coding gene of nCas9 protein, a coding gene of pyrimidine deaminase and a coding gene of adenosine deaminase, and a second expression plasmid containing gRNA, wherein the gRNA is a spacer sequence of a targeted target gene, and the length of the spacer sequence is 20 nt. The double-base editor can edit the base A and the base C at the same time, and an efficient and accurate tool basis is provided for the research fields of pichia pastoris-based recombinant protein evolution or chassis cell transformation and the like.

The invention relates to the technical field of lactobacillus rhamnosus, in particular to lactobacillus rhamnosus and application thereof in intervention of uric acid synthesis. The strain is lactobacillus rhamnosus LRa06 of which the preservation number is CGMCC (China General Microbiological Culture Collection Center) NO.33730. The strain has an inhibition effect on metabolic enzymes in a uric acid synthesis route, and especially has an inhibition effect on adenosine deaminase and xanthine oxidase. The strain inhibits uric acid synthesis and reduces serum urate by inhibiting metabolic enzymes of the two uric acid synthesis pathways, and has application prospects in preparation of uric acid reducing preparations.

An oligonucleotide is provided that can induce adenosine deaminase 1-specific editing activity. The oligonucleotide is used to induce site-specific editing of a target RNA, including: a first oligonucleotide that determines the target RNA; and a second oligonucleotide connected to the 5' side of the first oligonucleotide. The first oligonucleotide consists of: a target corresponding nucleotide residue corresponding to the adenosine residue in the target RNA; an oligonucleotide of 10 to 24 residues, which is connected to the 3' side of the target corresponding nucleotide residue and has a base sequence complementary to the target RNA; and an oligonucleotide of 3 to 6 residues, which is connected to the 5' side of the target corresponding nucleotide residue and has a base sequence complementary to the target RNA. The second oligonucleotide has a residue number of 2 to 10, forms a complementary chain with the target RNA, and the complementary chain contains at least one mismatched base.