32 results about "Golgi apparatus" patented technology

The invention discloses a near-infrared ratio type fluorescent probe for rapidly and reversibly detecting histamine (HA), a preparation method of the fluorescent probe and application of the fluorescent probe in dynamic cell imaging, and belongs to the technical field of fluorescence imaging and biosensing. The preparation method comprises the following specific steps: synthesizing aza-BODIPY molecules (1a-1d, 2a-2d) with different halogenations, and meanwhile, connecting the aza-BODIPY molecules with hydroxyl groups at 3 and 5 sites with a hydrophilic PEG (Polyethylene Glycol) fragment and halogen to obtain an HA probe HAPCyto with good water solubility; in addition, the hydroxyl aza-BODIPY molecules are continuously subjected to substitution, halogenation and hydrolysis reaction to obtain carboxyl-containing aza-BODIPY molecules, then the aza-BODIPY molecules are subjected to amidation reaction by utilizing the site, and different organic fragments are respectively connected to obtain a cell membrane targeted HA probe HAPPM, a golgi apparatus targeted HA probe HAPGA and a HaloTag HA fluorescent probe HAPHaloTag. When HA exists in a solution, the developed probe can realize rapid reversible detection of the near-infrared ratio of histamine, also shows good reversible property in mast cells, and can realize dynamic monitoring of histamine in cells.

The present invention is directed to a cargo-loaded cholesteryl ester nanoparticle with a hollow compartment (“cholestosome”) consisting essentially of at least one non-ionic cholesteryl ester and one or more encapsulated active molecules which cannot appreciably pass through an enterocyte membrane in the absence of said molecule being loaded into said cholestosome, the cholestosome having a neutral surface and having the ability to pass into enterocytes in the manner of orally absorbed nutrient lipids using cell pathways to reach the golgi apparatus. Pursuant to the present invention, the novel cargo loaded cholestosomes according to the present invention are capable of depositing active molecules within cells of a patient or subject and effecting therapy or diagnosis of the patient or subject.

The invention discloses application of brassica rapa L. polysaccharide targeted Golgi apparatus stress in preparation of vascular endothelial cell protection drugs, and belongs to the field of traditional Chinese medicine application. Pharmacological studies show that the Brassica rapa L. polysaccharide can reduce the expression of Golgi apparatus related proteins GRASP65 and p-GRASP65 and the ratio of p-GRASP65 / GRASP65, and the expression level of cell related proteins is reduced along with the increase of the concentration of the Brassica rapa L. polysaccharide. A transmission electron microscope is adopted to observe the form of the Golgi apparatus, and it is further proved that the Brassica rapa L. polysaccharide has the effect of inhibiting the stress response of the Golgi apparatus. Mechanism research shows that the Brassica rapa L. polysaccharide can inhibit Golgi apparatus stress and improve endothelial cell functions by regulating RvE1 / Ras / ERK signal axes. The invention provides an experimental basis for application and development of the Brassica rapa L. polysaccharide in treatment of vascular diseases.

The present invention related to the provision of genetically modified fungal cells, such as yeast cells with an improved ability for producing and secreting different recombinant proteins. The improved ability is obtained by disruption in intracellular transport between the Golgi and the endosome. In particular embodiments, the disruption is achieved by downregulation or deletion of the gene encoding a Tda3p homolog. The fungal cell and method of the invention would allow for large-scale production of recombinant proteins in fungal cells.

The invention provides a preparation method and application of a multifunctional targeting nano material for treating various cancers. A biotin derivative with a biotin receptor targeting capability is modified on the surface of polycyclodextrin, and is mixed and assembled with a Golgi apparatus targeting photosensitizer and a PD-L1 in-situ inhibitor, so that the constructed novel multifunctional targeting nano material can accurately recognize various tumor cells in a targeting manner, and the enrichment of the photosensitizer at a tumor part is increased; the accumulation in the golf apparatus can be further realized, and the structure and the function of the golf apparatus can be obviously damaged. In addition, the nano drug delivery system can also realize effective PD-L1 down-regulation, and the photodynamic synergistic immune anti-tumor effect is further enhanced. The novel multifunctional targeted nano drug-loading system prepared by the invention solves the problems of poor targeting and biological solubility and limited immune response ability and treatment effect of the body of the traditional photosensitizer, and provides an innovative strategy for realizing targeted photodynamic synergistic immunotherapy of cancers.

The present invention provides a gene for increasing glucose oxidase (GOD) production in Pichia pastoris and its use. Specifically, the present invention provides a method for constructing a glucose oxidase (GOD)-producing strain, wherein the activity of a vesicle secretion-promoting factor is enhanced or a vesicle secretion-promoting factor is introduced into the starting strain. Experiments have confirmed that co-expression of the VAMP4 gene, which is involved in the vesicle transport pathway from the Golgi apparatus to the plasma membrane, significantly increases extracellular GOD production in Pichia pastoris.

Disclosed herein include organelle complexes populations. The organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. In some embodiments, the organelle complexes are isolated or derived from floating cells and / or frozen cells. In some embodiments, the organelle complexes are isolated or derived from cells contacted with a surfactant at a concentration at or above the critical micellar concentration (CMC) for the surfactant. At least about 80% of the mitochondria of the organelle complexes are capable of maintaining structural integrity in an extracellular environment. Also provided herein are methods for generating first organelle complexes populations.

Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.

The invention relates to a Golgi apparatus targeting carbon dot nano-enzyme carrying GP73-siRNA. L-cysteine is used as a Golgi apparatus targeting raw material, a Mn-CDs nano-enzyme is prepared from L-cysteine, neutral red and manganese dioxide through a one-step solvothermal method, a PEI-HA complex is coupled to the surface of the Mn-CDs nano-enzyme to prepare a Mn-CDs-PEI-HA nano-enzyme carrier, and the Golgi apparatus targeting carbon dot nano-enzyme carrying GP73-siRNA is prepared through a one-step solvothermal method. And further carrying GP73-siRNA to construct a gene-loaded complex Mn-CDs-PEI-HA / GP73-siRNA, so as to target a liver cancer cell Golgi apparatus under the guidance of fluorescence / magnetic resonance bimodal imaging to carry out gene silencing treatment. The gene-loaded complex constructed by the invention has good fluorescence / magnetic resonance bimodal imaging capability, Golgi apparatus targeting capability and tumor hypoxic microenvironment improvement effect, so that integration of imaging and treatment is realized.

The present application provides a chimeric shuttle receptor comprising: (a) a heterologous binding moiety capable of specifically binding to a target protein, and (b) a polypeptide derived from a Golgi apparatus-related shuttle receptor; the chimeric shuttle receptor is used for reducing expression of a target protein. Methods of producing and using the chimeric shuttle receptors are also provided.

Disclosed herein include methods, compositions, and kits suitable for use in enhancing adoptive T cell therapy. In some embodiments, the method comprises contacting isolated organelle complexes with a population of T cells to generate a population of T cells comprising the organelle complexes. The organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. The population of T cells can exhibit one or more of enhanced expansion capability, enhanced cytotoxicity against target cells, enhanced resistance to exhaustion, and enhanced persistence, as compared to a population of T cells that do not comprise exogenous organelle complexes

The present invention provides a system of polynucleotides including: (a) a first polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a milk protein; (b) a second polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a first copy of a mammalian kinase; and (c) a third polynucleotide including a nucleic acid sequence encoding a Golgi sorting signal sequence and a second copy of said mammalian kinase, wherein the first polynucleotide, the second polynucleotide, and the third polynucleotide are operably linked. Further provided are cells and compositions including the system of polynucleotides, as well as methods of suing same, such as for producing a recombinant milk protein or a plurality thereof.

The invention discloses an application of a trimethylbenzofuran piperidinone derivative in sensitizing STING agonist antineoplastic treatment, and the derivative is (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidin-1-yl) ketone. The invention also discloses an application of the trimethylbenzofuran piperidinone derivative in sensitizing STING agonist antineoplastic treatment, and the derivative is (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidin-1-yl) ketone. In the prior art, through a strategy of knocking out STIM1 through CRISPR-Cas9, although cGAMP-mediated I-type interferon secretion is improved, intracellular calcium ion concentration is remarkably reduced due to irreversibility of knockout, calcium ion balance is destroyed, and processes of subsequent antigen cross presentation and the like are not facilitated. According to the present invention, the small molecule is developed to achieve the separation of STIM1 and STING, and the intracellular calcium ion steady state is maintained while the STING endoplasmic reticulum retention is released, such that the tumor treatment effect of the STING agonist is effectively enhanced, and the clinical immune response rate is expected to be significantly improved. Specifically, the invention provides a small molecule compound (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidine-1-yl) ketone, which has excellent biological activity, can promote the separation of STIM1 and STING and promote the transport of STING from endoplasmic reticulum to a Golgi apparatus, and obviously improves the level of an STING agonist secreting IFN-beta.

The invention belongs to the technical field of agriculture and biology, discloses application of arabidopsis thaliana GALT4 in regulation and control of pollen development, and verifies that the arabidopsis thaliana GALT4 gene is a plant male fertility key regulation and control gene with high pollen specificity expression, and the CDS sequence is shown as SEQ ID NO.1 for the first time. Polarity elongation of the pollen tube is maintained by regulating and controlling spatial and temporal distribution of methyl esterified pectin in the pollen tube. Knockout or silencing of the homologous gene in a plant can lead to reduction of pollen viability, abnormal development of pollen cell nucleus, malformation of pollen morphology, slowing of pollen germination rate and hindering of pollen tube polarity elongation. The invention provides a brand new molecular target for plant pollen morphological development and pollen tube growth regulation and control, can be used for fertility regulation and control and hybrid seed production optimization of cruciferous crops, and has important theoretical research value and industrial application prospect.

This invention discloses cotton GhGot1L This invention relates to genes and their application in creating heat-resistant cotton breeding materials, belonging to the field of cotton genetics and molecular breeding technology. This invention focuses on genetic analysis of the heat-resistant locus qPV-D05, which has been identified in the upland cotton genome. Through gene editing, it was determined that the causal gene at the qPV-D05 locus encodes a Golgi transporter protein. GhGot1L This invention corrects the problem through cloning experiments. GhGot1L Based on the reference genome annotation information, cloned GhGot1L The actual gene coding sequence was obtained, and molecular markers were developed for the heat-resistant genotypes generated by gene editing, which can be used to breed heat-resistant upland cotton germplasm.

The present invention discloses a fluorescent probe for dual detection of superoxide anions and peroxynitrosyl ions by targeting the Golgi apparatus, and its preparation method and application, belonging to the technical field of active oxygen probes. The fluorescent probe of the present invention has the molecular formula: C 42 H 54 F3N2O5S + The structural formula is: After the probe reacts with superoxide anions, the fluorescence intensity at 720 nm increases. Subsequently, after reacting with peroxynitrosyl ions, the fluorescence at 720 nm decreases and the fluorescence at 460 nm increases. The probe can also be used to detect superoxide anions and peroxynitrosyl ions in the Golgi apparatus of living cells, generating fluorescence images using confocal fluorescence microscopy. This suggests that the probe has potential practical value.

The invention relates to an engineering strain for intestinal delivery of a GLP-1 / GIP dual-receptor stimulant and application thereof, the strain takes uracil nutrition deletion type saccharomyces boulardii as a chassis strain, and a polypeptide expression module is integrated in a genome of the chassis strain; the polypeptide expression module comprises a secretory signal peptide coding sequence and a polypeptide coding sequence fused with the secretory signal peptide coding sequence, and the polypeptide coding sequence codes polypeptide formed by connecting a plurality of glucagon-like peptide-1 repetitive units and a plurality of glucose-dependent insulinotropic polypeptide repetitive units in series; restriction enzyme cutting sites which can be cut by yeast endogenous protease are arranged between the adjacent repeating units, so that the polypeptide is cut in a Golgi apparatus or a secretion processing process, and mature GLP-1 agonist and GIP agonist are released. The strain can be orally taken, can continuously control the delivery of GLP-1 / GIP dual agonists in an accurate dosage manner, so that the obesity and metabolic disorder are effectively improved, and the administration compliance and safety are improved.

The invention belongs to the technical field of fluorescent probes, and particularly relates to a Golgi apparatus targeting calcium ion fluorescent probe, a preparation method and application. The probe disclosed by the invention is simple to synthesize and relatively high in yield; according to the invention, specific and rapid detection of calcium ions in an aqueous solution is realized; according to the invention, the detection of calcium ions in the living cell Golgi apparatus is realized.

The invention belongs to the technical field of biology, and discloses a method for increasing the expression quantity of recombinant human serum albumin by optimizing a protein synthesis pathway, aiming at the problem of bottleneck in improving the efficiency of expressing recombinant human serum albumin by pichia pastoris in the prior art, the method comprises the following steps: S1, constructing an rHSA expression strain to obtain a candidate strain A / GS115; s2, carrying out multi-gene combination overexpression recombinant engineering bacterium construction by taking the candidate strain A / GS115 as a host bacterium; and S3, fermenting the constructed recombinant engineering bacteria to prepare the recombinant human serum albumin. According to the present invention, the related proteins RET2, BET3, YIP3, EMP24 and SEC22 in the protein synthesis pathway in pichia pastoris are subjected to overexpression, such that the transfer of the newly synthesized proteins from the endoplasmic reticulum to the Golgi apparatus can be easily mediated, and the newly synthesized proteins can be secreted to the extracellular region so as to improve the expression quantity of the rHSA;

The present specification relates to a composition for preventing or treating osteoarthritis, wherein the composition targets glycosylation. More specifically, the composition can inhibit abnormal accumulation of UDP-GlcNAc in the cytoplasm of chondrocytes and restore normal UDP-GlcNAc distribution to the endoplasmic reticulum and / or Golgi body, and thereby inhibit excessive O-GlcNAcization caused by cytoplasmic UDP-GlcNAc accumulation in chondrocytes, and can ultimately prevent or treat osteoarthritis.

The invention provides a copper delivery electrospun fiber scaffold targeting a Golgi apparatus as well as a preparation method and application of the copper delivery electrospun fiber scaffold, and belongs to the technical field of biological medicines. The invention provides a copper delivery system of a targeted Golgi apparatus for the first time, GHK-Cu is used as a stable copper source and continuously releases copper ions for cellular uptake, and LNPs is used for delivering mRNA (messenger ribonucleic acid) encoding a copper partner ATOX1. Upregulated ATOX1 transports copper ions to Golgi apparatus through ATP7A / B, thereby enhancing the activity of copper dependent proteins. In addition, ATOX1 promotes copper-dependent transport of ATP7A and Rac1 to plasma membrane, and synergistically accelerates new angiogenesis. According to the material system, copper accumulation in the golf ball body is remarkably increased, and the LOX activity is improved to 1.78 times that of a control group. The invention establishes a novel targeted Golgi apparatus copper delivery strategy, and provides a feasible treatment method for aplastic disorders (such as fascia defect) caused by insufficient copper-dependent protein activation.

A chimeric shuttling receptor, comprising: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from a Golgi-related shuttling receptor. The chimeric shuttling receptor is used for reducing the expression of the target protein. Also provided are methods for producing and using the chimeric shuttling receptor.

The invention provides a superoxide anion free radical activated indometacin-transretinoic acid prodrug, a preparation method of an albumin compound of the superoxide anion free radical activated indometacin-transretinoic acid prodrug and application of a composition prepared from the compound and an antitumor drug. The prodrug compound and an anti-tumor drug are prepared into the composition, the prodrug is activated by superoxide anion free radicals overexpressed in a tumor microenvironment, prototype drugs of indometacin and transretinoic acid are released, the damage effect of the transretinoic acid on a Golgi apparatus structure of a tumor cell is remarkably enhanced, and excellent broad-spectrum applicability is shown; and the device can be adapted to various drug delivery systems and disease models.

Disclosed herein include methods, compositions, and kits suitable for use in reducing or preventing oxidative stress and / or reductive stress. In some embodiments, the method comprises contacting a redox-sensitive composition with an effective amount of a disclosed redox-modulating composition. In some embodiments, the method comprises administering to a subject in need an effective amount of a disclosed redox-modulating composition. In some embodiments, the redox-modulating composition comprises isolated organelle complexes. Organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. In some embodiments, the redox-modulating composition comprises isolated mitochondria. The redox-modulating composition can be capable of reducing levels of ROS and / or RNS in a redox-sensitive composition and / or cells.

The application relates to application of nilotinib in preparation of anti-neuroblastoma drugs, and discloses a novel signal axis driving neuroblastoma progression: USP39-KLF1-SNARE-Golgi diffusion, and finds a new use of nilotinib in preparation of anti-neuroblastoma drugs. The application finds that nilotinib can effectively destroy the protein interaction between USP39 and KLF1. The destruction leads to the decrease of KLF1 protein stability, the inhibition of the transcription activity of KLF1, the down-regulation of the expression of VAMP3 / VAMP8, the weakening of the SNARE complex function, the reversal of the Golgi diffusion phenotype, and the slowing down of the growth of neuroblastoma. Nilotinib is a marketed drug, and has clear human safety data, so the clinical transformation can be accelerated.

This disclosure provides improved cell lines for manufacture of protein, considerably reducing the cost of commercial production. The cell lines are obtained by selecting cells from a mixed population for one or more characteristics that support protein production on a non-specific basis, such as the level of endoplasmic reticulum, Golgi apparatus, and / or other desired phenotypic features, compared with other cells in the starting mixture. Particularly effective producer cell lines can be obtained by preparing the cells for functional selection by making cell hybrids. A gene encoding a heterologous protein of interest may be transfected into the cells before or after one or more cycles of fusion and selection. Depending on the protein product being expressed, cell lines may be obtained that produce eight grams or more of protein per liter of culture fluid.

The invention provides a Golgi apparatus targeted nano-drug with a photodynamic and immunotherapy synergistic amplification effect and a preparation method thereof. The nano-drug is co-assembled by a hydrophobic photosensitizer and a high-molecular polymer DSPE-PEG2000-TK-IMC with a Golgi apparatus targeting effect through an intermolecular force. The photosensitizer forms an NIR-II absorption structure through J aggregation, has deep imaging and oxygen-independent photodynamic effects, and can efficiently generate active oxygen in an oxygen-deficient environment. ROS is induced by illumination to split TK bonds, so that directional release of immunomodulatory molecules is triggered, oxidative stress and immunogenic cell death are triggered in Golgi bodies, and meanwhile, a PGE2-mediated immunosuppression pathway is blocked. The system realizes synergistic enhancement of photodynamic therapy and immune activation, and has significant tumor microenvironment remodeling and precise treatment potentials.

The invention discloses a novel AIE photosensitizer as well as a preparation method and application thereof. The molecule takes triphenylamine as an electron-donating group and iodobenzoselenazole as an electron-withdrawing group, and is constructed through thiophene, benzothiadiazole or carbazole units. Researches show that DZTSe can efficiently generate type I and type II active oxygen, has low dark toxicity and good mitochondrial targeting ability, and is distributed in organelles such as endoplasmic reticulum, Golgi apparatus and the like. Under illumination, DZTSe induces pyroptosis by activating a caspase-1 / GSDMD pathway, releases DNA by damaging mitochondria, activates a cGAS-STING pathway, and promotes expression of type I interferon and inflammatory factors. Animal experiments show that DZTSe can significantly prolong the lifetime of mice with prostatic cancer, convert an immunosuppression microenvironment into an immune support state, and effectively inhibit primary and distal tumors.

The present invention related to the provision of genetically modified fungal cells, such as yeast cells with an improved ability for producing and secreting different recombinant proteins. The improved ability is obtained by disruption in intracellular transport between the Golgi and the endosome. In particular embodiments, the disruption is achieved by downregulation or deletion of the gene encoding a Tda3p homolog. The fungal cell and method of the invention would allow for large-scale production of recombinant proteins in fungal cells.

The present invention discloses a fluorescent probe, a preparation method thereof, and an application thereof, and belongs to the technical field of fluorescent probes. The preparation method of the fluorescent probe provided by the present invention comprises the following steps: the preparation method comprises stirring a compound I as shown in I and 6-methoxyquinoline in DMF, and separating and purifying the fluorescent probe compound. The fluorescence of the fluorescent probe synthesized by the present invention can be selectively quenched by chloride ions, and near physiological conditions, the Stern-Volmer constant for chloride ions is not affected by pH. The compound has good Golgi apparatus targeting ability, and its fluorescence intensity changes with the change of chloride ion concentration in the Golgi apparatus, proving that the probe can be used to detect changes in chloride ion concentration in the Golgi apparatus.