32results about How to "Improve expression level" patented technology

The application belongs to the technical field of biological medicine, and particularly relates to a kind of lipid nanoparticles and engineered immune cells. The application first provides an engineered immune cell, the engineered immune cell includes immune cell, lipid nanoparticle and exogenous drug;The exogenous drug is loaded in the lipid nanoparticle;When the immune cell is contacted with the lipid nanoparticle, the immune cell and the lipid nanoparticle endocytosis occur to obtain the engineered immune cell. In another aspect, the application also provides a kind of lipid nanoparticles that can improve the encapsulation efficiency of mRNA and the delivery efficiency of immune cells.

The present application relates to the field of cell therapy and regenerative medicine, and in particular to a kind of old age adipose tissue-derived mesenchymal stem cells pretreated by MK8722, its preparation method and application.The method comprises the following steps: the old age adipose tissue-derived mesenchymal stem cells are inoculated in culture solution containing MK8722 to carry out in vitro treatment, and first cells are obtained;The culture solution containing MK8722 is removed, and the first cells are washed to obtain second cells;Second cells are cultured using culture solution without MK8722, and the old age adipose tissue-derived mesenchymal stem cells pretreated by MK8722 are obtained.The cells obtained by the method, relative to the old age adipose tissue-derived mesenchymal stem cells without the in vitro treatment, show at least one improvement in proliferation capacity, continuous passage amplification capacity, adipogenic differentiation capacity or chondrogenic differentiation capacity, and can be used to prepare products for treating or alleviating osteoarthritis.

The invention relates to an RNA (Ribonucleic Acid) coding element and a method for carrying out gene editing on a plant by using the RNA coding element. In particular to an RNA (Ribonucleic Acid) coding element, an sgRNA bidirectional enhanced expression element, a carrier capable of carrying out efficient gene editing in a plant and a method for carrying out gene editing on the plant. The vector can improve the gene editing efficiency and the targeting specificity of the plant, and the sgRNA bidirectional enhanced expression sequence can further improve the gene editing efficiency and the targeting specificity of the plant.

The application discloses a method for culturing three-dimensional cell spheres of alveolar type II epithelial cells under a condition without exogenous matrix glue, and comprises the following steps: A, perfusion digestion treatment is performed on mouse lung tissue to obtain a single cell suspension; B, the single cell suspension is inoculated into a culture dish coated with anti-CD31 antibody and anti-CD45 antibody for immunoadhesion culture, under the culture condition, cells expressing CD31 and CD45 are subjected to immunoadhesion, meanwhile, fibroblasts with strong adhesion ability are attached to the surface of the culture dish, so that non-epithelial cells are removed, and unattached cells are collected to obtain an enriched ATII cell population; and C, the unattached cells are inoculated into a low-adhesion culture environment for suspension culture, and the culture system contains signal regulatory factors for maintaining the stability of the ATII cell phenotype, and the enrichment degree of epithelial cells is improved, and the interference of non-target cells is reduced through the combination of negative immunoadhesion of the anti-CD31 antibody and the anti-CD45 antibody and short-time adhesion treatment.

The application discloses a drought-inducible promoter of Cotinus coggygria and application thereof, a DNA sequence of the promoter is shown as SEQ ID No. 1, proCcDT1 is cloned from a promoter region of a CcDT1 gene of the Cotinus coggygria and is verified through experiments. First, elements of the promoter region are analyzed, and the result shows that the elements contain multiple cis-acting elements that can be involved in response to abiotic stress. Then, proCcDT1-GUS transgenic plants are treated through 10% PEG simulation drought, and it is found that GUS staining of the transgenic plants is enhanced, so it can be concluded that the GUS expression level driven by the promoter is increased under drought induction conditions, and it is also proved that the promoter can be induced by drought stress. The promoter proCcDT1 is introduced into an Arabidopsis genome through genetic transformation technology, so that the promoter can be stably induced by drought stress in a heterologous plant, and the practical application has important value for cultivating drought-resistant Cotinus coggygria varieties.

The application relates to an anti-apoptosis baculovirus expression vector, which realizes a broad-spectrum anti-apoptosis effect by expressing siRNA targeting a Sf-caspase-1 and Tn-caspase-1 common sequence of Spodoptera frugiperda and Trichoplusia ni insect cells through the vector. The baculovirus vector contains a specific DNA sequence, the DNA sequence contains an siRNA sequence targeting the Sf-caspase-1 and Tn-caspase-1 common sequence transcribed by an RNA polymerase III promoter; the recombinant virus of the vector can express double-stranded small RNA in a host cell, silences the caspase-1 encoded by the host cell through an RNA interference pathway, thereby inhibiting the apoptosis of the host cell, and significantly improving the expression level of an exogenous protein. The application can be used for industrialized production of protein preparations and vaccines.

The application belongs to the field of plant genetic engineering, and particularly relates to a series of expression vectors designed based on Capsicum annuum vein yellowing virus (PoPeVYV). The application proves that co-transfection of PoPeVYV aRNA and the expression vector can improve the expression efficiency of exogenous proteins in an infected object. The vector can not only efficiently express exogenous proteins in a host, but also realize assembly of virus-like particles. In summary, the application develops a new virus vector and a co-expression system thereof, and provides a new selection for a plant expression system.

The application relates to application of miRNA and / or a miRNA agonist in preparation of a medicine for preventing and / or treating myocardial ischemia-reperfusion injury, and belongs to the technical field of biological medicine. The medicine is administered through skeletal muscle injection, is secreted to a circulation system after expression on the skeletal muscle, is delivered to heart tissue by taking extracellular vesicles as carriers, and thus regulates myocardial cell functions. According to the animal model verification result, the miRNA can be delivered to the heart by the extracellular vesicles secreted by the skeletal muscle through injection of the medicine into the skeletal muscle, the expression level of the miRNA in the I / RI heart is significantly increased, the myocardial infarction area is effectively reduced, myocardial cell apoptosis is inhibited, myocardial fibrosis is reduced, and heart function is improved, a brand-new intervention strategy is provided for treatment of myocardial ischemia-reperfusion injury and related heart diseases, and a brand-new idea is opened up for research and development of heart failure treatment medicines.

This invention relates to a therapeutic target for metabolic dysfunction-related fatty liver disease (MAFLD) and hepatitis, and its application, belonging to the field of biomedical technology. This invention is the first to discover that the expression level of DPF2 in Lgr5-positive hepatocytes can specifically determine the severity of MAFLD and MASH. Based on this finding, this invention significantly improves the pathological structure of MASH by overexpressing DPF2 in Lgr5-positive hepatocytes, demonstrating that targeting Lgr5-positive hepatocytes in the liver can be a therapeutic strategy for metabolic-related fatty liver disease. This invention provides a novel therapeutic target for the treatment of MAFLD / MASH, which has good specificity and broad application prospects.

The invention discloses a lactobacillus rhamnosus CQFP202457 and application of the lactobacillus rhamnosus CQFP202457 and ellagic acid in relieving exercise fatigue, and belongs to the technical field of microorganisms. The preservation number of the lactobacillus rhamnosus CQFP202457 is CGMCC (China General Microbiological Culture Collection Center) No. 29784. The composition provided by the invention contains ellagic acid and lactobacillus rhamnosus CQFP202457, and can significantly prolong the exercise exhaustion time, reduce the content of fatigue-related metabolites in serum, improve the total antioxidant ability of serum, liver and skeletal muscle and the antioxidant enzyme gene expression level, and improve the exercise-induced liver and skeletal muscle tissue damage. The strain and the composition thereof provided by the invention can be used as exercise synbiotics for preparing products for relieving exercise-induced fatigue, improving exercise performance, reducing exercise-induced oxidative stress or relieving exercise-induced muscle injury.

The invention relates to the technical field of gene engineering, in particular to a Cilp gene enhancer and application thereof. Through integration and verification of an ATAC-seq map, a single cell transcriptome data set and multi-omics data, the Cilp gene enhancer is obtained through screening, and a mouse inner ear microinjection system is utilized to further prove that the Cilp gene enhancer can significantly enhance the transcriptional activity of an EGFP protein gene in cochlea type 2 neuronal cells. Compared with other sequences with similar sizes, the Cilp gene enhancer provided by the invention can drive specific expression of a reporter gene or a Cilp gene in cochlear neurons, especially type 2 neurons, and the gene expression level is remarkably improved. The Cilp gene enhancer provided by the invention is suitable for biological materials such as recombinant DNA (deoxyribonucleic acid), vectors or adenoviruses, can be used for promoting transcription of EGFP (enhanced green fluorescent protein) genes or other genes in cochlea neurons, and provides a new targeting tool for gene therapy of hereditary hearing loss.

The present application relates to the field of biomedical technology, and more particularly to a sorting method of a mesenchymal stem cell subpopulation secreting high hepatocyte growth factor and a screening method of a sorting index. The application of cell surface protein CD10 as an index in sorting the mesenchymal stem cell subpopulation secreting high hepatocyte growth factor. The present application successfully associates CD10 with HGF secretion function and strong repair efficiency for the first time, providing an unprecedented method capable of directly and non-destructively screening and enriching a high therapeutic potential cell subpopulation from a heterogeneous cell population. The method overcomes the core bottleneck of traditional mesenchymal stem cell products, which are not uniform in function and unstable in efficacy, making it possible to sort based on the CD10 surface marker, thereby enabling the preparation of a mesenchymal stem cell subpopulation product with higher HGF expression level, more significant repair function and better quality uniformity. It has important clinical application value and industrialization prospect.

The invention discloses a targeted modified fungal luciferase protein and application thereof in enhancing the bioluminescence intensity of fungi or plants. The invention aims to solve the problems of subcellular localization disorder and activity loss caused by deletion of natural membrane localization signals of fungal luciferase in heterologous systems such as plants and the like. The invention creatively provides an innovative positioning reconstruction strategy: removing a natural membrane positioning sequence at the N end of the Luz protein, and then carrying out double modification on the N end and the C end. The method specifically comprises the following steps: fusing endoplasmic reticulum targeting signal peptide at the N end to guide a target protein into an endoplasmic reticulum cavity; meanwhile, endoplasmic reticulum retention signal peptide is fused at the C end of the Luz protein, so that the Luz protein is stably retained in an endoplasmic reticulum, subcellular localization of the Luz protein is accurately reconstructed, Luz is ensured to be localized into cytoplasm, and the Luz protein is prevented from entering a secretion path, so that the expression level and activity of the Luz protein in a fungus or plant system are remarkably improved, and the bioluminescence intensity of fungi or plants is remarkably enhanced.

The present invention relates to a novel sequence peptide having hair growth promotion and hair loss prevention activity. The peptide has an activity that can promote or induce cell proliferation or activity related to hair formation, and thus a composition comprising the same can be useful in preventing hair loss or promoting hair growth.

This invention relates to a heparin hydrolase mutant and its recombinant expression method, belonging to the field of enzyme engineering technology. This invention heterologously expresses a heparin hydrolase derived from *Chromobacterium amazonense* in *Escherichia coli*, and modifies the heparin hydrolase by adding a TrxA solubilizing tag and performing site-directed mutagenesis on certain amino acid sites. The mutant G37T / N145D / V281K exhibits a catalytic efficiency 1.96 times higher than the original strain, playing an important role in the application and industrial production of heparin hydrolase.

The application belongs to the field of biology and particularly relates to a VP3 protein mutant recombinant Seneca virus strain and construction and application thereof. It is found for the first time that the lysine at the 82nd position of the VP3 protein of the Seneca virus is a key amino acid for regulating translation arrest, and after the site is mutated to alanine, the translation inhibition function of the VP3 protein is significantly weakened; further, through a molecular cloning technique, an infectious clone of the VP3 protein at the 82nd site is successfully constructed, and a corresponding recombinant virus strain rSVA VP3-K82A is rescued; compared with the wild type SVA, the virus proliferation ability of the recombinant virus strain is decreased, the virus titer is reduced, the immune escape ability is reduced, and the influence on the translation function of the host is weakened; not only does it provide a key tool for in-depth analysis of the functional mechanism of the VP3 protein in the virus replication cycle and immune escape, but also serves as a candidate strain of attenuated live vaccine, and provides a new idea for the development of Seneca virus prevention and control strategies.

The present application relates to the field of molecular biology, and particularly relates to application of a reagent for regulating NDRG2 expression level in preparation of a preparation for treating preeclampsia. 6 A methylation modification level; and reducing the expression level of NDRG2. 6 A methylation modification regulates NDRG2 mRNA stability and further promotes the molecular mechanism of NDRG2 expression. 6 A methylation modification regulates NDRG2 mRNA stability and further promotes the molecular mechanism of NDRG2 expression. Based on this mechanism, by regulating the expression level of NDRG2, the present application effectively inhibits the invasion, migration and proliferation of placental trophoblast cells, promotes the expression of apoptosis and inflammatory factors, which provides a new technical means for targeted treatment of preeclampsia.

The application discloses a genetically engineered bacterium with high ergothioneine yield and a construction method and application thereof. The genetically engineered bacterium is a recombinant microorganism with the following characteristics or changes in a host for ergothioneine production: (1) overexpression of betaine homocysteine methyltransferase BHMT; (2) overexpression of ergothioneine synthase EgtE, EgtD and Egt1; and (3) knockout of spermidine synthase gene spec and tRNA methyltransferase gene trmB in the genome. In the fermentation production of ergothioneine, the genetically engineered bacterium is supplemented with betaine, and the addition of methionine is reduced. The utilization rate of methionine reaches more than 70%, and the yield of ergothioneine reaches more than 10 g / L, thereby providing a new scheme with improved economy for the industrial production of ergothioneine.

The invention provides a squalene production strain capable of automatically cracking and application of the squalene production strain in preparation of squalene. The strain is obtained by taking saccharomyces cerevisiae as an original strain and enhancing the activity of a gene encoding the following enzyme, the enzyme comprises acetyl coenzyme A transferase, 3-methyl-3-hydroxyglutaryl coenzyme A synthase, 3-hydroxy-3-methylglutaryl coenzyme A reductase, mevalonate kinase, mevalonic acid-5-phosphate kinase, mevalonic acid-5-pyrophosphate decarboxylase, isopentenyl pyrophosphate isomerase, farnesyl pyrophosphate synthase and squalene synthase. The invention further discloses a preparation method of the enzyme. After the engineering bacterium finishes efficient synthesis of squalene, cells are cracked to efficiently release squalene accumulated in the cells into fermentation liquor, so that separation and purification of downstream products are more convenient, downstream processes are simplified, the cost is reduced, and the production economy is integrally improved.

This invention discloses a sheep small RNA miR-433-3p and its applications, belonging to the field of animal molecular breeding and reproductive biology technology. The mature nucleotide sequence of miR-433-3p is shown in SEQ ID NO.1. Studies have shown that after transfection of miR-433-3p mimic into sheep follicular granulosa cells, the expression level of miR-433-3p significantly increased, the cellular EdU positivity rate decreased, the cell cycle G0 / G1 phase ratio increased while the S and G2 phase ratios decreased, the apoptosis rate increased, and the expression of proliferation-related genes / proteins and apoptosis-related genes / proteins changed accordingly; transfection of miR-433-3p inhibitor showed the opposite changes. Based on the above results, miR-433-3p can be used to regulate the proliferation and apoptosis of sheep follicular granulosa cells, and can serve as a candidate molecular marker for constructing an evaluation system related to sheep high fertility screening / breeding, providing experimental basis for the subsequent establishment of molecular evaluation indicators for fertility; its mimics, inhibitors, expression vectors or kits containing it can also be used to screen candidate nutritional factors, bioactive factors or reproductive regulatory factors that affect granulosa cell function.

The application belongs to the technical field of bioengineering, and discloses a screening platform for optimizing the interval of baculovirus bidirectional promoters, a screening method and application. The application constructs a high-resolution screening platform with a reference gene integrated in a virus skeleton, and through two-stage screening, first, a transcription enhancement interval is determined by rough screening in the 98-245bp interval, and then, fine screening is performed in the transcription enhancement interval by 1bp steps, so that the optimal interval length of the p10 and polh promoters is determined as 153bp by normalizing the reference fluorescent signal to correct errors such as infection efficiency. The interval is applied to a recombinant adeno-associated virus (rAAV) packaging system, transcription interference between the bidirectional promoters is eliminated, the expression balance of Rep and Cap proteins is restored, the rAAV genome replication efficiency is improved by 3 times, the empty shell rate is reduced from 84.9% to 20.1%, and the transduction efficiency in mammalian cells can be maintained.

This invention belongs to the field of regenerative medicine technology, specifically relating to bone marrow-derived macrophages expressing CD301b, TIMD4, and LYVE-1, their preparation method, and applications. The steps are as follows: Primary mouse bone marrow mononuclear precursor cells are cultured in expansion medium for 7 days to obtain macrophages, with the medium changed every 2-3 days during this period; on the 7th day, the expansion medium is replaced with a high-efficiency induction medium and cultured for 46-50 hours to obtain bone marrow-derived macrophages expressing CD301b, TIMD4, and LYVE-1. This invention, through a time-series strategy of TGF-β pre-induction combined with IL-4 / IL-5 / IL-13 synergistic induction, can significantly enhance the expression levels of CD301b, TIMD4, and LYVE-1 in BMDM, making their function close to that of natural cardiac resident macrophages, and effectively exerting anti-inflammatory, repair-promoting, and ventricular remodeling-inhibiting effects after AMI.

The application belongs to the technical field of biology and particularly relates to application of MYBS3 gene in regulating content of anthocyanin and luteolin in honeysuckle flower. The nucleotide sequence of the MYBS3 gene is shown as SEQ ID NO. 1, and the MYBS3 gene is up-regulated in expression under low-temperature stress by spraying Ca 2+ in honeysuckle flower. The recombinant plasmid containing the MYBS3 gene can obviously improve expression levels of anthocyanin synthase (ANS) and flavone synthase (FNS) in leaves. The recombinant plasmid containing the MYBS3 gene can obviously improve content of anthocyanin and luteolin in leaves of honeysuckle flower.

The application discloses a traditional Chinese medicine composition for treating Parkinson's disease and a traditional Chinese medicine preparation and application thereof. The traditional Chinese medicine composition comprises, in terms of weight parts, 9-20 parts of Schisandra chinensis, 10-30 parts of Ophiopogon japonicus, 3-15 parts of Radix ginseng rubra, 10-30 parts of Uncaria, 10-30 parts of Cuscuta chinensis, 10-30 parts of Semen ecliptae prostratae, 3-5 parts of ginger and 5-15 parts of Jujube. The application further discloses a preparation of the traditional Chinese medicine composition. The traditional Chinese medicine composition and the preparation thereof can effectively improve the motor function of Parkinson's disease patients, reduce the dosage of Madopa, and thus reduce the related side effects caused by dopamine replacement therapy.

The application discloses a culture medium for CAR-T cell culture and application thereof, and relates to the fields of immunology, molecular biology and cell engineering. The culture medium comprises a basic culture medium and dihydroeugenol. It is found in research that by adding dihydroeugenol in the culture medium, the proportion of central memory T cells of CAR-T cells can be effectively improved, and the expression levels of PD1 and LAG3 can be reduced. In addition, dihydroeugenol can also promote the secretion of IL2, TNF and INFgamma, and enhance the killing function of CAR-T cells. Experiments show that the ability of D-CAR-T to secrete IL2, TNF and INFgamma is significantly higher than that of ordinary CAR-T, and D-CAR-T also has a higher T CM cell proportion and a lower exhaustion level after performing the killing function.

The present invention relates to the technical field of recombinant protein design and synthesis, and specifically discloses a recombinant spider silk protein and its synthesis method. The recombinant spider silk protein provided by the present invention undergoes optimized design such as replacement of the N-terminal and C-terminal domains of different spider species and different spider silk types, and large fragment recombination of the middle functional domain repeat sequences. A series of recombinant spider silk proteins are obtained through cell-free protein synthesis methods and prokaryotic fermentation expression. Compared with natural spider dragline silk proteins or other existing recombinant spider silk proteins, they generally have good composite mechanical properties and high-temperature resistance characteristics. At the same time, they have a relatively small molecular weight, significantly improving the total expression level and soluble expression, providing new raw materials and design and synthesis methods for the development of high-performance biomaterials.

The application provides a SAA recombinant protein and a preparation method and application thereof, and the preparation method comprises the following steps: S10, constructing a recombinant plasmid: a gene sequence of an amino acid sequence shown in SEQ ID NO: 1 or a nucleotide sequence corresponding to an amino acid sequence shown in SEQ ID NO: 2 is inserted into a pET-28a(+) plasmid which is double-digested by NdeI and XhoI after being double-digested by NdeI and XhoI, so that a recombinant plasmid is obtained; S20, transforming and culturing the recombinant plasmid; S30, protein expression and purification. The application solves the technical problem that the stability of the currently recombinantly expressed SAA protein in vitro is poor, and achieves the technical effect of improving the stability of the SAA recombinant protein in vitro.

A method for collecting blood and preparing serum from giant freshwater prawns includes: feeding the prawns a nutrient-rich diet under low-temperature conditions, collecting blood from their second walking legs, and obtaining purified serum through blood coagulation, serum precipitation, and further processing; the nutrient diet includes at least a protocatechuic acid-chitosan oligosaccharide derivative, which is obtained by grafting protocatechuic acid onto chitosan oligosaccharide. This invention preserves the life of individual giant freshwater prawns during blood collection and increases the amount of blood and serum collected.

The invention provides a DNA tetrahedron drug compound for treating retinal ischemia reperfusion injury, and belongs to the field of eye disease treatment and molecular biology, and the tetrahedron drug compound is connected with YAP saRNA. The tetrahedral drug compound can efficiently and safely deliver YAP saRNA, improve the expression level of YAP in cells and inhibit oxidative stress and ferroptosis of retinal cells, so that RI / RI is effectively treated, and a new choice is provided for treating the tetrahedral drug compound.

The invention belongs to the technical field of zearalenone induced liver injury, and particularly relates to application of a Keap1-Nrf2-quercetin conjugate in repairing zearalenone induced pig liver injury. The invention discloses the Keap1-Nrf2-quercetin conjugate for the first time, and the Keap1-Nrf2-quercetin conjugate can be used for remarkably improving the expression levels of cell antioxidant related proteins NQO1, Keap1, HO-1 and Nrf2, successfully activating the expression of Nrf2 and downstream genes thereof, improving the antioxidant capacity of cells and relieving the damage of ZEA to the cells; the ZEA can inhibit response of a Keap1-Nrf2 signal channel to ROS (reactive oxygen species) while triggering oxidative stress of cells, and the change is relieved by introducing a Keap1-Nrf2 quercetin conjugate. The invention provides the application of the Keap1-Nrf2-quercetin conjugate in the ZEA-induced pig liver injury, and a new intervention target is provided for developing treatment of novel ZEA-induced intestinal injury.