203 results about "Cytidine" patented technology

The instant specification provides for evolved base editors which overcome deficiencies of those in art (including increased efficiency and / or decreased requirement for specific sequence-context at an editing site) and which are obtained a result of a phage-assisted continuous evolution (PACE) system. In particular, the instant specification provides for evolved cytidine base editors (e.g., based on APOBEC1, CDA, or AID cytidine deaminase domains) which overcome deficiencies of those in art (including increased efficiency and / or decreased requirement for specific sequence-context at an editing site) and which are obtained a result of a phage-assisted continuous evolution (PACE) system.

The invention relates to a metabolic marker for diagnosing diabetic secondary osteoporosis and application of the metabolic marker, and belongs to the technical field of biological medicine. The method comprises the following steps: firstly, screening out obviously different metabolites between osteoporosis (DOP) and osteoporosis-free groups (DM) in diabetic patients; and secondly, screening out significant difference metabolites between the primary osteoporosis patient and the healthy control group, and excluding the intersection of the two groups of difference metabolites to obtain metabolites which exclude the influence of the primary osteoporosis. Metabolic markers, namely cytosine nucleotide, ketoglutaric acid and triethylamine, are obtained through further screening, and when the three metabolic markers are independently used, AUCgt; the AUC during combined use is 0.99, so that the limitation that the sensitivity of the traditional bone mineral density detection on the osteoporosis recognition in the diabetic population is insufficient is effectively made up.

The invention discloses an Escherichia coli mutant strain which is classified and named as Escherichia coli, the strain number is CR-TK01, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.35554. The invention further discloses a preparation method of the Escherichia coli mutant strain. The Escherichia coli CR-TK01 with high yield of cytidine provided by the invention has the characteristics of high yield and stable high-yield character, glucose is used as a carbon source, the Escherichia coli CR-TK01 is used for fermentation production of cytidine under a certain fermentation condition, and the yield of cytidine is as high as 97.23 g / L; the strain is not degraded after passage for more than 20 generations, the performance of high-yield cytidine is maintained in the passage fermentation process, and the yield of cytidine in passage fermentation is improved. The method for producing cytidine through fermentation is simple in process, high in yield and suitable for industrialization.

The present disclosure relates to recombinant microorganisms, such as bacteria, e.g., E. coli, engineered to have improved flux and / or yield of the DXP pathway. Cells can be engineered to comprise nucleotide sequences comprising dxs polypeptides (EC 2.2.1.7); dxr polypeptides (EC 1.1.1.267); ispD polypeptides (EC 2.7.7.60); ispE polypeptides (EC 2.7.1.148); ispF polypeptides (EC 4.6.1.12); ispG polypeptides (EC 1.17.7.1 or EC 1.17.7.3); ispH polypeptides (EC 1.17.7.4); and idi polypeptides (EC 5.3.3.2). Cells can be further engineered to express redox polypeptides, e.g., ferredoxins, flavodoxins, and / or flavodoxin / ferredoxin--NADP reductases (EC 1.19.1.1 or EC 1.18.1.2), e.g., to increase the activity of the ispG and / or ispH polypeptides. Some or all of the nucleotide sequences can be heterologous to the cells. The recombinant microorganisms can be used in methods of isoprenoid production.

The invention relates to the field of anti-tumor drugs, and discloses an anti-diffuse large B-cell lymphoma pharmaceutical composition, a preparation method and application thereof, the composition comprises azacitidine and Sanilisoma, the composition can improve the accumulation amount of VDUP1 protein in a cell nucleus through the combination of the methylation effect of DNMT3b on VDUP1 protein and the inhibition of XPO1 activity by Sanilisoma, and the anti-diffuse large B-cell lymphoma pharmaceutical composition can be used for preparing anti-diffuse large B-cell lymphoma. The chemotherapy effect is remarkably improved, the sensitivity of the DLBCL to the Sanisole is remarkably improved, and particularly, the pharmaceutical composition has a remarkable curative effect on R / R DLBCL patients.

The invention discloses escherichia coli recombinant engineering bacteria for producing citicoline as well as a construction method and application of the escherichia coli recombinant engineering bacteria, and belongs to the technical field of biology. A genome cytidine deaminase coding gene cdd is knocked out on the basis of Escherichia coli K-12 MG1655, a temperature control expression vector PBV220 is used for expressing a SpnCCT-SpnCKI-pRpLpyrG mutant gene cluster, the expression of choline transporter protein BetT is enhanced, the gene locus of Escherichia coli ptsH-ptsI-crr is destroyed, meanwhile, a glucose permease gene glf and a glucokinase gene glk from Zymomonas mobilis are introduced, and the mutant gene cluster of the Escherichia coli K-12 MG1655 is obtained. The escherichia coli recombinant engineering bacterium ZMCB02NCL capable of efficiently biosynthesizing citicoline is obtained, and the preservation number of the escherichia coli recombinant engineering bacterium ZMCB02NCL is CCTCC NO: M 2025483.

The invention discloses a method for improving the survival rate of probiotics based on cell membrane regulation and control. The method comprises the following steps: adding a lipid component in a probiotic culture process and / or adding the lipid component as a protective agent after probiotic culture, the lipid component added in the probiotic culture process comprises cytidine diphosphate diacylglycerol and / or stearic acid; the lipid component added as the protective agent after culture comprises cardiolipin and / or oleic acid. The method breaks through the research category of a traditional optimized freeze drying process (such as a protective agent and parameters), and key lipid molecules directly related to resistance are screened out through a lipidomics means and establishment of a screening method of stress resistance related lipid molecules; through a dual mode of supplementing key lipid components and precursor substances thereof, the tolerance of the strain is improved from the metabolic level, and the freeze-drying resistance and the survival rate of the strain are remarkably improved.

The invention provides inter alia methods of treating cancer comprising administering to a subject a cytidine triphosphate synthase 1 (CTPS1) inhibitor and a B-cell lymphoma 2 (BCL2) inhibitor.

The present invention belongs to the technical field of genetic engineering. Provided are a single-base editor, a deaminase used therein, and the use thereof. The technical problems to be solved are to identify a naturally occurring cytosine deaminase without sequence preference, construct a base editor, and improve the efficiency and scope of base editing. In order to solve the technical problems above, a cytosine base editor is provided. The cytosine base editor is a fusion protein, wherein the fusion protein is a protein containing a cytidine deaminase, a Cas protein and a uracil-DNA glycosylase inhibitor, and the cytidine deaminase is a protein having an amino acid sequence of positions 28-164 of SEQ ID NO. 2. Further provided is the use of the fusion protein above and a biomaterial related thereto in plant single-base editing. The single-base editor can improve the efficiency of cytosine base editing and accurately mediate the base mutation of a target, and is widely applicable in the cells of maize and even other plants.

The invention belongs to the technical field of bioengineering, and particularly relates to a phosphorylcholine cytidyltransferase mutant and application thereof. On the basis of phosphorylcholine cytidyltransferase MsCCT001, any one or more of N194D, K91E, G98S, S148A and C156V are mutated respectively, a series of phosphorylcholine cytidyltransferase mutants are obtained through screening, and the obtained mutants have the advantages that in the reaction of catalyzing phosphorylcholine and cytidine triphosphate to generate citicoline, the yield of the obtained mutants is increased, and the yield of the obtained mutants is increased. The catalytic efficiency of the compound is 1.90-2.41 times that of initial phosphorylcholine cytidyltransferase, and a biological enzyme preparation with higher catalytic capacity is provided for preparing citicoline by a biological conversion method.

The invention belongs to the field of synthetic biology and metabolic engineering, and particularly relates to high-yield sialyl acyl-N-tetrasaccharide a escherichia coli as well as a construction method and application thereof. According to the method, recombinant escherichia coli is taken as an initial strain, and a beta1, 3-galactosyl transferase gene wbgO from escherichia coli O55: H7 is integrated into a chromosome in a multi-copy form, so that a lactyl-N-tetrasaccharide efficient synthesis chassis strain without plasmid dependence is constructed. And overexpression neuC, neuB, neuA and Nm3ST are introduced through plasmids, so that the supply of glycosyl donors is improved. The expression intensity of uridine diphosphate-N-acetylglucosamine synthesis pathway genes glmM and glmUS is regulated through ribosome binding site engineering, and a cytidine triphosphate regeneration module is introduced, so that the yield of sialyloyl-N-tetrasaccharide a is increased. And finally, the maximum titers of the sialyloyl-N-tetrasaccharide a respectively reach 2.238 g / L and 8.178 g / L by shaking a flask and culturing in batches. The invention provides an efficient technical scheme for industrial production of complex breast milk oligosaccharide.

The invention relates to a T7RNA polymerase-guided bacillus subtilis cytosine base editor and application thereof, and belongs to the technical field of gene editing. A T7RNAP guided cytosine base editor is developed in bacillus subtilis, four kinds of cytidine deaminase from different sources are selected to be fused with T7RNAP, the cytidine deaminase is fused at the N end of the T7RNAP, and a fusion body with the highest mutation activity is obtained through screening; the cytosine base editor is used for evolving a global transcriptional regulatory factor CodY, in-situ evolution of genome protein is achieved, and the expression level of bovine-derived beta-lactoglobulin (beta-Lg) is improved. According to the cytosine base editor, the operable range of the cytosine base editor on a genome is widened, multi-site base (C-T) editing can be realized, and an editing window is large.

The application discloses cytosine deaminase and related biological materials and applications thereof, and belongs to the technical field of proteins. The technical problem to be solved by the application is to mine natural sequence-unpreferred cytosine deaminase, and to improve base editing efficiency and range. The application provides a protein with amino acid sequence of SEQ ID NO. 2 at positions 15-143. The application also provides application of the above protein and related biological materials thereof in single base editing. The application uses macro-genome big data, performs comprehensive and systematic bioinformatics mining analysis on novel cytosine deaminase, and selects a protein L85 highly similar to known DddA protein structure but with low sequence similarity for function verification. The protein L85 has high catalytic activity as cytosine deaminase, and has no target sequence preference restriction, and can be widely applied to in-vitro and in-cell / tissue gene editing, and has good application potential and development value.

This invention provides a simple synthesis method for Fe-Ag / CMP nanopolymers with catalase-like activity and their application in hydrogen peroxide detection, belonging to the field of bioanalytical technology. The method of this invention includes the following steps: (1) preparing a 4-hydroxyethylpiperazine thiosulfonic acid (HEPES) standard buffer solution; (2) adding cytidine monophosphate (CMP) to the solution obtained in step (1) and stirring until the CMP is fully dissolved. Then, under vigorous stirring, ferric nitrate aqueous solution and silver nitrate aqueous solution are added sequentially, at which point the colorless solution turns pale yellow; (3) transferring the mixed solution obtained in step (2) to a centrifuge tube, centrifuging at 12000 rpm for 8 min to collect the precipitate, washing it three times with distilled water, and finally adding 4 mL of distilled water to obtain a 1 mg / mL Fe-Ag / CMP suspension. This invention utilizes CMP as a biological ligand and Ag... + Fe 3+ Simultaneous coordination, and the reducing amino groups on CMP can directly transfer Ag... + In situ reduction to silver nanoparticles (AgNPs) led to the one-step synthesis of a bimetallic Fe-Ag / CMP nanopolymer with hydrogen peroxide-like nanozyme activity, enabling the detection of hydrogen peroxide.

Aspects of the disclosure relate to mRNAs comprising a relatively low abundance of cytidine: adenosine (CA) dinucleotides that benefit from increased stability relative to mRNAs containing more CpA dinucleotides. The disclosure also relates to methods of modifying an mRNA sequence to improve stability. In some aspects, the disclosure relates to mRNAs comprising modified mRNA sequences with relatively reduced numbers of CpA dinucleotides, and compositions comprising mRNAs with relatively reduced numbers of CpA dinucleotides.

This invention discloses a method for directed evolution of antibiotic-resistant Escherichia coli strains based on cytidine deaminase, belonging to the field of microbial directed evolution and genetic engineering technology. This method uses E. coli as the host, introducing a recombinant plasmid expressing an optimized double-stranded cytidine deaminase mutant. Utilizing the low toxicity and high mutagenicity of this mutant, continuous passage evolution is carried out under gradient concentrations of aminoglycoside antibiotics (kanamycin and streptomycin). Combined with whole-genome sequencing, molecular docking, and reverse genetics verification, the A145T missense mutation in the wcaE gene is identified as the core functional site. This invention overcomes the shortcomings of traditional spontaneous and chemical mutagenesis, which suffer from low efficiency and significant strain damage. The mutation type is controllable, the evolutionary cycle is short, and the obtained engineered strains can tolerate up to 300 mg / L kanamycin while exhibiting streptomycin cross-resistance, and the genetic stability of the tolerance trait is strong. This method is simple, highly reproducible, and suitable for industrial breeding of stress-resistant E. coli, and can be widely applied in antibiotic fermentation, industrial microbial culture, and other scenarios.

The invention belongs to the technical field of genetic engineering and microbial engineering, and particularly relates to a method for improving the production level of cytidine and application. According to the recombinant escherichia coli, a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd are subjected to overexpression. Specifically, the fusion expression of a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd is carried out, and the fusion expression is carried out by adopting Linkers with different properties. During application, the recombinant escherichia coli can be used for producing the cytidine by fermentation, so that the production level of the cytidine is improved. According to the invention, recombinant Escherichia coli which is over-expressed and fused with a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd is constructed, and the recombinant Escherichia coli can be used for efficiently fermenting and producing cytidine.

The protein complex comprises cytidine deaminase and helicase. In some embodiments, the cytidine deaminase is an altered cytidine deaminase. In some embodiments, the protein complexes convert 5 methylcytosine to thymine. Kits, compositions, and methods of use comprising the protein complexes of cytidine deaminase and helicase are also described.

The present disclosure relates to immunogenic compositions comprising at least one Yersinia pestis (Y. pestis) antigen, an aluminum salt adjuvant, and an oligonucleotide comprising an unmethylated cytidine-phospho-guanosine (CpG) motif. The immunogenic compositions are suitable for stimulating an immune response against Y. pestis in a subject in need thereof. The present disclosure also relates to kits and methods using the immunogenic compositions, or two separate compositions which together comprise the antigen, the aluminum salt adjuvant, and the oligonucleotide.

The invention discloses a method for improving the water-resistant straw-like dwarf virus capability of rice based on D14 protein site editing and application of the method, according to the scheme, an interaction interface of RGSV P3 and rice D14 protein is analyzed through a structural biology means, and it is determined that the 102nd part, with the D14 binding site, of D14 and P3 is aspartic acid (Asp, D). Then, accurate editing of the D14 gene in the rice is realized by utilizing a cytidine base editor (CBE) system, and the site is mutated into asparagine (Asn, N), so that D14 (D102N) transgenic rice is obtained. Disease resistance identification confirms that the mutant has significant resistance to RGSV. Furthermore, a homozygous non-transgenic disease-resistant material which does not contain exogenous transgenic ingredients is obtained through genetic screening and has a good breeding application prospect.

The invention belongs to the technical field of medicine synthesis, and particularly relates to a cytidine precursor compound as well as a preparation method and application thereof. The cytidine precursor compound provided by the invention is a compound with a structure as shown in a formula (I) or a pharmaceutically acceptable salt thereof, in the formula (I), R2 is selected from one of hydrogen, formyl, acetyl, propionyl, n-butyryl and isobutyryl; r1, R3, R4, R5 and R6 are respectively and independently selected from two or three of hydrogen, hydroxyl and fluorine. The cytidine precursor compound provided by the invention is convenient to prepare oral and intravenous injection preparations, can be effectively converted into a raw drug in vivo, has bioavailability and tumor aggregation which are obviously higher than those of a cytidine raw drug, has an excellent drug treatment effect, has obvious clinical advantages, and greatly meets the clinical application requirements of cytidine antitumor drugs.

The present disclosure relates generally to pharmaceutical dosage forms comprising azacitidine and chidaridine.

The present invention relates to a method for producing α-sialyl-β-D-galactoside saccharides, particularly α-sialyl-(2→3)-β-D-galactoside saccharides and α-sialyl-(2→6)-β-D-galactoside saccharides, from a β-D-galactoside saccharide, a sialic acid donor, and an enzyme with β-galactoside α-sialyltransferase activity. The enzymes with β-galactoside α-sialyltransferase activity used herein do not exhibit catalytic activity towards the hydrolysis of cytidine 5'-monophospho-N-acetyl-neuraminic acid to cytidine and N-acetyl-neuraminic acid. The method can be performed in vitro and in vivo using a genetically engineered cell comprising a nucleic acid encoding said enzyme. Further, said process may be adapted to produce the sialic acid donor CMP-Neu5Ac from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, a cytidine phosphate (CMP, CDP or CTP) and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), optional a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3).

The invention discloses an application of N4-acetylcytidine (ac4C) in preparation of a medicine for delaying senescence and prolonging life. The method is realized by reducing the overall ac4C modification level of RNA (including rRNA, tRNA, mRNA, ncRNA and the like) in a living body, and specifically can be realized by inhibiting the expression of a key catalytic adapter protein THUMPD1 or using an inhibitor targeting THUMPD1 / ac4C. The invention also provides an application of a reagent for detecting the aging degree or diagnosing aging-related diseases, such as Alzheimer's disease, mitochondrial diseases, tumors and the like, in preparation of a detection kit based on the ac4C modification level of a specific target RNA molecule, and provides a brand new target spot and strategy for aging intervention and accompanying disease diagnosis and treatment.

The present invention provides a composition comprising a cytidine analog, in particular decitabine or azacitidine. In particular, the present invention provides a composition comprising a cytidine analog, in particular decitabine or azacitidine, which is useful for topical application. The present invention also provides the use of such compositions for medical conditions in the keratinizing and non-keratinizing epithelium / skin, such as the treatment of human papillomavirus (HPV)-related pre-cancerous conditions.

The present invention relates to a photosensitive RNA cytidine base editor and use thereof. The photosensitive RNA cytidine base editor can cause deamination of cytidine (C) in a target double-stranded RNA to uridine (U), thereby achieving editing of C to U. The photosensitive RNA cytidine base editor comprises: (1) a dCas-eADAR2dd N-pMagHigh fusion protein, and (2) an nMagHigh-eADAR2dd C fusion protein.

The present disclosure generally relates to the use of linear nucleic acid primers for the amplification of a target nucleic acid sequence, for example, in a cell-free environment. In some embodiments, compositions of the linear nucleic acid primers are provided. For example, in some embodiments, the linear nucleic acid primers comprise a guanosine or a cytidine at 3′ terminal end. In some embodiments, the linear nucleic acid primers have been optimized to prevent primer-homodimer and / or hairpin formation and to exclude cumbersome codon sequences. In some embodiments, methods are provided for the amplification of a DNA template fragment using the linear nucleic acid primers. Thus, in some cases, the use of the nucleic acid primers, as described herein, may allow for the reduction in amplification of non-specific hybridization events while allowing for the amplification of the target nucleic acid sequence.

The invention discloses a composite functional bacterial agent for regulating nucleoside substances in Luzhou-flavor liquor as well as a preparation method and application of the composite functional bacterial agent, and belongs to the technical field of liquor production. The compound functional bacterial agent is prepared from a liquid culture of pichia kudriavzevii GTY837 and a liquid culture of pichia kudriavzevii PKW194, or a liquid culture of pichia kudriavzevii GTY837 and a liquid culture of pichia kudriavzevii PKW194, or a liquid culture of pichia kudriavzevii PKW194, or a liquid culture of pichia kudriavzevii GTY837 and a liquid culture of pichia kudriavzevii PKW194. According to the composite functional bacterial agent provided by the invention, through the synergistic effect of the GTY837 and the PKW194, the content of seven nucleoside substances such as cytidine and hypoxanthine in the Luzhou-flavor liquor is effectively increased. The microbial agent solves the technical problem of unstable generation of nucleoside substances in traditional fermentation, and realizes synergistic improvement of the flavor and health attribute of Baijiu by enriching nucleoside substances with the functions of dispelling the effects of alcohol, protecting nerves and the like while enhancing the mellow feeling, delicate flavor and other sensory qualities of the Baijiu.

The invention relates to combination and combination therapies, particularly for the treatment of a myeloid malignancy, such as acute myeloid leukemia (AML) or a myelodysplastic syndrome (MDS). The combination therapies include (i) a recombinant binding protein comprising (a) an ankyrin repeat domain specifically binding CD3 and (b) at least one, at least two, or at least three further ankyrin repeat domains, wherein each of the at least one, at least two, or at least three further ankyrin repeat domains specifically binds one tumor-associated antigen, suitably one selected from the group consisting of CD123, CD33 and CD70, and (ii) a Bcl-2 inhibitor, e.g., venetoclax, or a pharmaceutically acceptable salt thereof and / or (iii) a hypomethylating agent, e.g., azacitidine or decitabine.