373 results about "Messenger ribonucleic acid" patented technology

A neurotrophic factor production promoting device is provided, which is able to promote production of a neurotrophic factor or neurotrophic factor-like substance in an affected area by a simple technique that, regardless of the place of treatment, can be performed without transplantation of cells or injection into the affected area, in order to prevent or treat various diseases such as brain diseases. In order to apply a high frequency alternating magnetic field in the range of 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700 MHz to 1000 MHz to cells at a magnetic flux density of no more than 0.01 Tesla, the neurotrophic factor production promoting device includes a high frequency electromagnetic wave generating means generating a high frequency electromagnetic wave of the abovementioned frequency, in which the magnetic stimulation by the high frequency alternating electromagnetic field of the abovementioned high frequency allows the intracellular concentration of calcium ions to be increased so that exocytosis of the neurotrophic factor group is induced, and the magnetic stimulation allows messenger ribonucleic acid (mRNA) of the neurotrophic factor group to be increased in the cells so that the synthesis and extracellular release of the neurotrophic factor group are promoted.

The invention provides an SNP (Single Nucleotide Polymorphism) molecular marker related with a back fat thickness trait of a pig and a detection method of the SNP molecular marker. By detecting exon regions of MARK4 (Microtubule Affinity-Regulating Kinase) genes of different pig species, one SNP locus (G-A) is found to exist at 55th basic group (mRNA (Messenger Ribonucleic Acid) total-length 2581th base group) in 3 minute UTR (Untranslated Region) of the MARK4 gene, an allele A is positively correlated with low back fat trait, and an allele G is positively correlated with high back fat trait. The invention provides the method for detecting the SNP molecular marker. Pig genomic DNA (Deoxyribonucleic Acid) to be detected is amplified by using a primer pair with the nucleotide sequence shown in SEQ ID NO.1-2 and is subjected to enzyme digestion by using pstI; if an enzyme-digested product is 251bp, a base group at a mutation is A; if the enzyme-digested product is 218bp, a base group at the mutation is G. According to the method provided by the invention, the back fat thickness of the pig can be predicted early, quickly and effectively at low cost; the SNP molecular marker can be used as a useful molecular marker for genetic improvement of the pig species.

The disclosure features methods of reducing or inhibiting an anti-drug antibody response in a subject, as well as methods of reducing or inhibiting unwanted immune cell activation in a subject to be treated with a messenger RNA (mRNA), comprising administering to the subject a mRNA, e.g., a chemically modified messenger RNA (mmRNA), encoding a polypeptide of interest, wherein the mRNA comprises at least one microRNA (miR) binding site for a miR expressed in immune cells, such as miR-126 binding site and/or miR-142 binding site, such that an anti-drug antibody response to the polypeptide or interest, or unwanted immune cell activation (e.g., B cell activation, cytokine secretion), is reduced or inhibited in the subject. The disclosure further provides therapeutic treatment regimens designed to reduce or inhibit AD A or unwanted immune cell activation (e.g., B cell activation, cytokine secretion) in a subject being treated with mRNA-based therapeutics.

The invention discloses a method for establishing an immunodeficiency mouse model. The method comprises the following steps of: (1) constructing DNA (deoxyribonucleic acid) which enables functions of an immune related gene in a mouse cell to be lost; (2) purifying mRNA (messenger ribonucleic acid), and cryopreserving in a super-purified water of RNAase free; (4) feeding a pregnant female mouse and a pseudopregnant female mouse under an SPF (specific pathogen free) level feeding condition; (5) taking a fallopian tube from the abdominal cavity of the pregnant female mouse in the step (4), and collecting fertilized ovum; (6) injecting the mRNA obtained in the step (3) into the pronucleus of the fertilized ovum in the step (5) by using a microinjection method; (7) transplanting the fertilized ovum obtained in the step (6) into the body of the pseudopregnant female mouse obtained in the step (4), and enabling the pseudopregnant female mouse to culture the second generation; (8) establishing a stable deficient mouse strain. The method has the advantages that a mouse model which completely does not have immunological rejection to allotransplantation can be obtained for testing.

The invention provides a micro-droplet type PCR (polymerase chain reaction) chip and a manufacture method thereof. The upper layer of the chip is a PDMS (polydimethylsiloxane) imaging cover plate formed by connecting a DNA (deoxyribonucleic acid) target fragment with a primer injection port, a single phase drop forming microstructure, a PCR reaction enzyme and an mRNA (messenger ribonucleic acid) substrate injection port, an oil phase reagent injection port, a drop straight line channel, a cross hybrid channel, a micro drop buffer channel, a micro drop PCR reaction chamber and a micro drop PCR waste liquid chamber, the lower layer of the chip is a glass substrate, and the upper layer and the lower layer are bonded and packaged to form a complete micro drop PCR chip. The manufacture method of the chip comprises the steps of firstly, manufacturing a mask plate; secondly, molding; thirdly, pouring a die; and fourthly, taking a figure. The PDMS figure at the upper layer of the chip adopts a multilayer filter structure, and the formed micro drop has a uniform size, stable grain diameter and strong controllability. Compared with traditional PCR chips, the micro drop PCR chip manufactured by the method has great advantages in hybrid effect, reaction efficiency, and high throughput.

The present invention relates to pharmaceutical compositions comprising a compound and a pharmaceutically acceptable carrier. The present invention is also directed to a method of treating a genetic disease caused by premature termination codons, or other conditions that render messenger ribonucleic acid (mRNA) susceptible to nonsense mediated RNA decay, in a subject. Also disclosed is a method of inhibiting nonsense mediated RNA decay and/or induction of autophagy. The present invention also relates to a method of identifying inhibitors of nonsense mediated RNA decay and/or inducing autophagy. The present invention further relates to a method of inhibiting nonsense mediated RNA decay and/or induction of autophagy in a subject.

The invention belongs to the field of pharmacy, and relates to application of a hydrogen sulfide donor to the preparation of a medicine for treating central nervous system disease, in particular to application of the hydrogen sulfide donor, namely sodium hydrosulfide, or allyl cysteine and analogues thereof to the preparation of the medicine for treating the central nervous system disease. Integral animal model experiments prove that the sodium hydrosulfide and propargyl cysteine can reduce the learning and memory impairment of rats, increase the content of hydrogen sulfide in hippocampus tissues of the rats, inhibit the messenger ribonucleic acid (mRNA) expression of a tumor necrosis factor alpha and a receptor I thereof, inhibit an inflammatory mediator in the hippocampus tissues of rats with dementia and inhibit inflammation-related enzymes in the hippocampus tissues of the rats, and can be used as a medicine for treating Alzheimer disease and other inflammation-related central nervous system degenerative diseases such as Parkinson disease.

The requirement of T cell activation for efficient expression of genes after messenger ribonucleic acid (mRNA) transfection is leveraged to identify and enrich antigen-specific T cells responding to antigen-pulsed dendritic cells (DCs). RNA transfection of marker genes is used for the selection and enrichment of antigen-specific T cells for use in adoptive immunotherapy. RNA-modified T cells are also used for the generation of enhanced effector populations for use in adoptive immunotherapy. Genes whose transient expression may significantly enhance the in vivo function of T cells (i.e., migratory receptors, anti-apoptotic genes or cytokines enhancing T cell proliferation/differentiation) are used in this modality.

The invention belongs to the field of application of feed additive, and relates to a porcine heat stress resistance selenium-rich composite bacteria feed additive and application thereof. Saccharomyces cerevisiae, lactobacillus acidophilus and streptococcus thermophilus are selected as strains, inoculated in a proper fermentation broth containing sodium selenite, transformation of selenium and probiotics growth accelerating agent in a volume ratio of 1:2:2, and fermented for 30 to 40 hours at a temperature of between 20 and 40 DEG C, throughput of 0.05m<3>/h, revolution of 70r/min and tank pressure of 0.04Mpa; and the selenium enriched composite bacteria preparation can be prepared through proper subsequent treatment. The daily gain and feed-weight ratio of a pig can be remarkably improved in a high-temperature environment by feeding the preparation, the oxidation resistance and immunity functions can be enhanced, the mRNA (messenger ribonucleic acid) level of heat shock proteins Hsp70 and Hsp27 can be reduced, and the effect of porcine heat stress resistance can be effectively played.

The invention discloses recombinant oral protein TAT-GH of tilapia, a preparation method for the recombinant oral protein TAT-GH and application of the recombinant oral protein TAT-GH. The method comprises the following steps of: extracting total messenger ribonucleic acid (mRNA) from a pituitary gland of male tilapia, performing reverse transcription-polymerase chain reaction (RT-PCR) to obtain complementary deoxyribonucleic acid (cDNA), performing PCR amplification to obtain a growth hormone (GH) gene of the tilapia, and amplifying a TAT-GH gene by a PCR overlap extension method; constructing a recombinant plasmid pET-32a(+)-TAT-GH; and transforming Escherichia coli BL21 (DE3) by using the recombinant plasmid pET-32a(+)-TAT-GH, efficiently expressing fusion protein TAT-GH in the form of an inclusion body under the induction of isopropyl thiogalactoside (IPTG), and purifying and renaturing to obtain active protein. The protein can effectively promote the growth and development of fries of the tilapia after being fed to the fries of the tilapia.

The invention discloses a molecular marker for diagnosing osteoporosis. The molecular marker is an EPS8L3 gene. The molecular marker has the advantages that as proved by research, the mRNA (messenger ribonucleic acid) expression level of EPS8L3 genes in the blood of osteoporosis patients is obviously reduced as compared with normal persons; reagent kits for diagnosing the osteoporosis can be prepared according to correlation between the EPS8L3 genes and the osteoporosis and can be clinically widely applied.

The invention discloses a kit for jointly detecting breast cancer 21 genes. A PCR (polymerase chain reaction) amplification primer consists of 21 primer pairs, and can be used for jointly detecting 21 gene-related expression quantities: Ki67, STK15, Survivin, CCNB1, MYBL2, MMP11, CTSL2, GRB7, HER2, GSTM1, CD68, BAG1, ER, PGR, BCL2, SCUBE2, ACTB, GAPDH, PRLPO, GUS, and TFRC. Output fragments of the 21 pairs of primers related in the invention are smaller than 100bp, so that PCR efficiency after reverse transcription of mRNA (messenger ribonucleic acid) extracted by a paraffin specimen is greatly improved, all primers of 21 genes are premixed, and time and labor for designing and synthesizing 21 pairs of genes by an experimenter are saved; the corresponding primers of the 21 genes are not needed to be added for premixing in mother liquor, and therefore, errors of reaction holes are greatly reduced. The kit disclosed by the invention is simple to operate, and capable of detecting on a quantitative PCR instrument by only adding a template, so that detection experimental time is greatly shortened. The kit disclosed by the invention is applicable to a single sample and multiple samplers, so that results can be quickly obtained, and therefore, scientific research and clinical detection requirements can be satisfied very well.

The invention finds a bioinformatics method according to gene sequence expression and gene prediction algorithms, wherein the method can be used for directly predicting and quantifying a long noncoding RNA (ribonucleic acid) and directly locking the LncRNA for further experimental verification. The method disclosed by the invention mainly comprises the following process: step one, collecting all the full-length mRNA (messenger ribonucleic acid) sequence data of a person; step two, removing the mRNA sequence comprising an extron of a coding protein; step three, forming a retrievable database by using LncRNA more than 200bp, and; step four, searching the existing gene expression sequence analysis data to identify over-expressed LncRNA from the analysis data; and step five, carrying out experimental verification. Thus, the over-expressed LncRNA in a specific cell tissue is predicted finally..

A method for probing a target sequence of messenger ribonucleic acid molecules (mRNA's) in a fixed, permeabilized cell, said target sequence including at least 30 non-overlapping probe binding regions of 15-100 nucleotides, comprising immersing said cell in an excess of at least 30 nucleic acid hybridization probes, each singly labeled with the same fluorescent label and each containing a nucleic acid sequence that is complementary to a different probe binding region of said target sequence; washing said fixed cell to remove unbound probes; and detecting fluorescence from said probes.

The invention aims to provide alpha amylase and an aspergillus niger strain for expressing the same. An amino acid sequence of alpha amylase is SEQ ID NO:1. A coding mRNA (messenger ribonucleic acid) sequence of alpha amylase is SEQ ID NO:2. The mutant strain aspergillus niger 1-B5 obtained by an ultraviolet mutagenesis method can efficiently express alpha amylase AclaP11 derived from aspergillus clavatus, with strain number of CGMCC (China General Microbiological Culture Collection Center) No.8443. The enzyme activity of aspergillus niger 1-B5 is 1.97 times that of an original strain. The optimum pH value and optimum temperature of alpha amylase AclaP11 expressed by aspergillus niger 1-B5 are respectively 6.0 and 50 DEG C. Compared with the original strain, aspergillus niger 1-B5 has the advantage that the enzymatic properties are not changed. Besides, aspergillus niger 1-B5 is a food safety fungus (GRAS(generally recognized as safe)), so that alpha amylase expressed by aspergillus niger 1-B5 in a secretory manner can be applied to the field of food additives, thus further broadening the market space of alpha amylase.

The invention provides a carbohydrate composition with a wound healing promoting effect and application of the carbohydrate composition. The carbohydrate composition contains at least five types of carbohydrate including carboxymethyl chitosan, rhizoma bletillae polysaccharides, fucoidan from fucus vesiculosus, heparan sulfate, chondroitin sulfate, oligosaccharides of hyaluronan and gulose aldehyde acid oligosaccharides. The carbohydrate composition and the application have the advantages that VEGFA (vascular endothelial growth factor A), FGF2 (fibroblast growth factor 2) and VE-cadherin protein mRNA (messenger ribonucleic acid) expression can be obviously improved by the carbohydrate composition, VEGF (vascular endothelial growth factor) secretion and skin fibroblast (HSF) and human immortalized keratinocyte (HaCaT) proliferation can be obviously promoted by the carbohydrate composition, and the obvious wound healing promoting effect can be realized by the carbohydrate composition; raw materials required by the carbohydrate composition are easily available, composition methods for the carbohydrate composition are simple, the carbohydrate composition is low in cost, easy to industrialize and wide in application range, and the like.

The invention relates to a novel method for culturing a multipotential stem cell without serum and a feeder layer. The method is stable, reliable, and can culture the multipotential stem cell for a long time and also can maintain the self-renewing of the multipotential stem cell. The method is characterized in that a culture system with clear chemical components and without serum and the feeder layer is adopted to perform suspension culture on the multipotential stem cell. According to the morphology of cultured stem cells after 15 generations, mRNA (Messenger Ribonucleic Acid) and protein expression of multipotential marker molecule, and analysis on cell chromatin karyotype, such novel generation system can maintain the properties of the multipotential stem cell during long-term culture. In addition, the differentiative potential of the multipotentail stem cell is further verified by 15-generation suspension culture in an in-vitro embryoid body formation experiment and an in-vivo teratoma formation experiment.

The invention relates to an mRNA (messenger ribonucleic acid) quick-extraction kit and a preparation method thereof. The kit comprises a separation device for solid-liquid separation, wherein the separation device at least comprises a layer of cellulose solid-phase carrier. The kit also comprises at least one tube containing a cell or tissue lysis solution, at least one tube containing a biotinylated Oligo (dT) probe, at least one tube containing a combination buffer solution, and at least one tube containing an elution buffer solution. The kit is simple, quick and efficient to operate, and is suitable for separating and extracting mRNA in a biological sample. The invention is mainly characterized in that the separation device uses cellulose as the solid-phase carrier, a fusion protein CBD-SA is used as a biological bridging agent, and a biologically-specific coupling method is used instead of the traditional physical adsorption method and chemical bonding method to activate the cellulose solid-phase carrier, so that the activated solid-phase carrier can be efficiently combined with the biotinylated reagent to be used for separating protein or nucleic acid.

The invention provides a goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and an application thereof. According to the goose-origin gene RIG-I and the application of the goose-origin gene RIG-I, mRNA (Messenger Ribonucleic Acid) sequences of the goose-origin gene RIG-I are amplified based on a duck-origin gene RIG-I, and the homology of the goose-origin gene RIG-I and the duck-origin gene RIG-I is found to reach 93.8%; heterogenously-expressed gRIG-I is proved to be capable of expressing an anti-NDV (Newcastle disease virus) effect after over-expressing transfection cells of the full-length gRIG-I (gRIG-I full) and a gRIG-I CARD structural domain (gRIG-I CARD); the gRIG-ImRNA levels are detected by respectively infecting primary-culture geese embryo fibroblasts (GEF) and 2-week Yangzhou geese by utilizing three NDV viruses; the gRIG-ImRNA levels are found to averagely rise after the GEFs are infected by the NDV; and after being infected by the NDV, the gRIG-ImRNA levels in tissues of lungs and air sacs of the geese averagely rise, and the virus growths in in-vitro and in-vivo experiments are all restrained.

The invention discloses a common mussel C type lysozyme and a preparation method thereof. The amino acid sequence of the common mussel C type lysozyme is shown by a sequence table SEQ ID No.1. The preparation method of the common mussel C type lysozyme comprises the following steps of: extracting common mussel gross RNA (ribonucleic acid), purifying mRNA (Messenger ribonucleic acid), preparing a cDNA (complementary deoxyribonucleic acid) template solution, cloning gene, building recombinant plasmid, expressing expression plasmid and purifying protein to obtain the C type lysozyme. The common mussel C type lysozyme is a new C type lysozyme which has strong inhibitory activity on various gram positive and negative microorganisms, and has application value in the aspects of developing food preservatives, feed additive, etc.

The invention discloses a bicistronic mRNA (messenger ribonucleic acid) expression vector which is suitable for cells of mammals and can realize high-efficient expression of exogenous genes and an application thereof. The bicistronic mRNA expression vector comprises connecting elements in the 5'-3' direction, namely (1) an open reading frame 1: containing the sequences of an early promoter/enhancer and an introne of cytomegalovirus, multiple clone sites and the sequence of a bovine growth hormone transcription terminator, which are sequentially arranged; (2) an open reading frame 2: containing the sequences of the early promoter/enhancer and the introne of the cytomegalovirus, the multiple clone sites and the sequences of the bovine growth hormone transcription terminator and a simian virus promoter, which are sequentially arranged; and (3) screening marker genes and the sequence of a simian virus 40 (SV40) terminator. The bicistronic mRNA expression vector has the high-efficient promoter, effective transcription termination signals, screening and gene amplification markers and two open reading frames, and can improve the expression efficiency of the exogenous genes and reduce the production cost.