34 results about "Cellular compartment" patented technology

The invention discloses a protein subcellular localization method and system based on gating multi-mode routing, and relates to the technical field of protein subcellular localization, and the method comprises the steps: obtaining a protein image and a protein sequence; the method comprises the following steps: respectively adopting a ConvNeXt-V2 network and an ESM-2 model to carry out feature extraction on a protein image and a protein sequence to obtain image features and sequence features, and then based on a Sparsemax gating network, obtaining multi-modal fusion features; according to the STRING database and the subcellular compartment distance matrix, a weighted adjacency matrix is constructed, a protein map is obtained, then in combination with multi-modal fusion features, through a positioning perception neural network, aggregated neighbor features are obtained, and in combination with geometric prototype decoding and hierarchical constraint loss, multi-label prediction is carried out, and an optimized prediction result is obtained. According to the method, the capability of capturing complex protein features is improved, the problem of insufficient single-modal information is solved, and the accuracy of positioning prediction is improved.

The present invention belongs to the field of microbial metabolic engineering and synthetic biology technology applications, and particularly relates to a Pichia pastoris engineered strain that produces high sclareol production, as well as its construction method and application. A sclareol biosynthesis pathway is constructed in a host strain, and the intracellular mevalonate metabolic pathway and the central metabolic pathway are optimized, thereby obtaining an engineered bacterial strain A; the host strain is Pichia pastoris; or, metabolic regulatory factors are overexpressed or knocked out in the above-obtained engineered bacterial strain A, thereby obtaining an engineered bacterial strain B; or, in the above-obtained engineered bacterial strain B, a cell compartmentalization strategy is used to target the synthesis pathway to the peroxisome and optimize it, thereby obtaining an engineered bacterial strain C. The present invention provides a Pichia pastoris chassis cell for synthesizing the diterpenoid compound sclareol and an engineered strain for synthesizing sclareol; by introducing and optimizing the expression of the sclareol synthesis pathway, the sclareol yields in shake flask batch fermentation and bioreactor batch fed-batch fermentation reach 631.6 mg / L and 10.5 g / L, respectively.

The invention relates to a gene delivery system for cytoplasm specific expression of PPM1K as well as a preparation method and application of the gene delivery system, and belongs to the technical field of preparation of gene delivery systems. The gene delivery system is preferably an adeno-associated virus (AAV), and the preparation method of the gene delivery system specifically comprises the following steps: inserting a PPM1K gene or a functional variant thereof into a vector, and adding a detectable label at a C terminal to form a recombinant plasmid; transforming the recombinant plasmids into competent escherichia coli, amplifying, and screening and extracting plasmids to obtain recombinant AAV plasmids; co-transfecting the recombinant AAV plasmid, the helper plasmid and the capsid plasmid into packaging cells by using a transfection reagent, collecting the cells and culturing supernate after transfection, and adding a freeze thawing buffer solution for repeated freeze thawing to obtain a cell lysis solution; centrifuging the cell lysis solution, taking supernate, purifying and filtering. According to the invention, the specific expression of the cell compartment of the therapeutic gene is realized through the vector design, and the obvious therapeutic effect and the safety advantage are shown.

The present invention relates to a multi-compartment microfluidic device comprising a first cellular compartment (10) within which a target organ is reproduced, a second cellular compartment (20), located below the first cellular compartment (10) and separated from it by a porous membrane (30), and a third cellular compartment (40) connected to the first cellular compartment (10) by a means forming a biological interface (50). Figure 1

Disclosed herein are related aspects of microneedle arrays comprising metal-organic framework (MOF)-vaccine biocomplexes and methods of using the same. The microneedle array also includes a dissolvable material that dissolves upon insertion into the subject's skin, thereby releasing the MOF-vaccine biocomplex. MOF dissolved in an acidic environment can be selected, so that targeted delivery of the vaccine to a specific cell compartment is realized. The invention also discloses a method for preparing the MOF-vaccine biological complex and the microneedle array.

The present invention relates to a multi-compartment microfluidic device comprising a first cellular compartment (10) in which a target organ of the digestive system is reproduced and a second cellular compartment (40) comprising neurons (400), the second cellular compartment (40) being connected to the first cellular compartment (10) by a means forming a biological interface (50) to allow communication via neuronal connection (401) between the first cellular compartment (10) and the second cellular compartment (40). Figure 1

The present application relates to a kind of cytoplasm-specific expression PPM1K gene delivery system and its preparation method and application, belong to gene delivery system preparation technical field.The gene delivery system is preferably adeno-associated virus (AAV), and its preparation method specifically includes the following steps: PPM1K gene or its functional variant is inserted into vector, detectable label is added at C-terminal, and recombination plasmid is formed;Recombinant plasmid is transformed into competent e. coli and is amplified, after plasmid is screened, recombination AAV plasmid is obtained;Recombinant AAV plasmid, auxiliary plasmid and capsid plasmid are co-transfected into packaging cell using transfection reagent, after transfection, cell and culture supernatant are collected, freeze-thaw buffer is added repeatedly freeze-thaw, and cell lysate is obtained;After centrifugation of cell lysate, supernatant is taken, and purified filtration is carried out.The present application realizes the cell compartment specificity expression of treatment gene by vector design, and shows significant treatment effect and safety advantage.

Aspects of the present application provide methods for analyzing biological samples. The methods can include flowing a biological sample through a fluidic channel, wherein individual cells from the biological sample can be compartmentalized. In certain aspects, at least 105 cells from the biological sample can be compartmentalized. At least a subset of the compartmentalized cells can be lysed to release nucleic acids, and the nucleic acids can be captured and / or analyzed.

To provide methods of reacting a cell nucleus informational molecule with an analytical biomolecule.SOLUTION: The invention relates to a system and methods of enhancing access to nuclear informational molecules, such as DNA, RNA, and proteins, by analytical biomolecules, such as transposome complexes, by treating nuclei with a nuclear permeability enhancer, and to methods of using nuclear membranes, cell membranes, and external compartmentalization approaches as contiguity preserving elements. The present invention provides, for example, a method of reacting a cell nucleus informational molecule with an analytical biomolecule, comprising contacting the cell nucleus with a nuclear permeability enhancer and reacting the informational molecule with the analytical biomolecule.SELECTED DRAWING: None

Methods of treating heart disease by modulating the cellular localization of the beta isoform calcium / calmodulin dependent protein kinase II delta (CaMKIIδ) via the disruption of post-transcriptional phosphorylation.

Constructs for detection of d-2-hydroxyglutarate (d-2-HG), and their use in determining the IDH1 / 2 mutational status of a biological sample obtained from a subject, monitoring a change in D-2-HG levels in a subject, and analysing D-2-HG in a biological sample obtained from a subject, comprising detecting for D-2-HG in the sample or subcellular compartment therein, and methods for the same. Nucleic acid molecules, vectors, cells, and pharmaceutical compositions are also described.

The invention discloses a saccharomyces cerevisiae engineering bacterium for efficiently synthesizing mogroside V based on cytoplasm and peroxisome double-cell-region indoor and a construction method of the saccharomyces cerevisiae engineering bacterium. Wherein the construction method comprises the following steps: integrating acetoacetyl coenzyme A thiolase ERG10, 3-hydroxy-3-methylglutaryl coenzyme A synthetase ERG13, truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase tHMG1, phosphomevalonate kinase ERG8, mevalonate pyrophosphate decarboxylase ERG19, pentenyl pyrophosphate isomerase IDI1 and geranyl pyrophosphate synthase ERG20 at a site of a saccharomyces cerevisiae genome GAL80, and carrying out enzymolysis, so as to obtain a saccharomyces cerevisiae strain; a squalene synthase ERG9, an epoxy squalene cyclization enzyme ERG1 and the like; aBC efflux protein PDR11, isopentene phosphokinase AtIPK, geranyl pyrophosphate synthase ERG20, endogenous squalene synthetase ERG9, epoxy squalene cyclization enzyme ERG1 and the like are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The yield of MG-V synthesized by shake flask fermentation of the saccharomyces cerevisiae engineering bacteria constructed by the invention reaches 117.6 mg / L, and the saccharomyces cerevisiae engineering bacteria have the advantages of high yield, strong metabolic flux directionality, wide industrial application prospect and the like.

Constructs for detection of d-2-hydroxyglutarate (d-2-HG), and their use in determining the IDH1 / 2 mutational status of a biological sample obtained from a subject, monitoring a change in D-2-HG levels in a subject, and analysing D-2-HG in a biological sample obtained from a subject, comprising detecting for D-2-HG in the sample or subcellular compartment therein, and methods for the same. Nucleic acid molecules, vectors, cells, and pharmaceutical compositions are also described.

A culture system for human dermal and epidermal layers of the skin is provided, which culture reproduces the histomorphology of native human skin. The system is highly reproducible and maintains the cellular diversity of human skin for extended periods of time, both in the types of cells that are present, and in the architecture of the tissue. Features of the culture include self-production of molecules and proteins required for maintenance and maturation of the diverse cellular compartments.

Hie disclosure provides methods for cell-based screening of genetically encoded protein binders, involving (a) providing recombinant eukaryotic cells that have a first nucleic acid encoding a membrane protein, and / or a secreted protein; and a second nucleic acid encoding a first fusion protein comprising one or more recombinant protein target-binding candidate fused to (A) a direct cellular compartment retention tag, or (B) an indirect cellular compartment retention tag; wherein the protein target-binding candidate, when expressed, is retained within a cellular compartment by means of the cellular compartment retention tag; (b) culturing the recombinant eukaryotic cell under conditions suitable to express the recombinant protein target and the protein target-binding candidate in the recombinant eukaryotic cell; and (c)detecting localization of the protein target in the recombinant eukaryotic cell in one or more of the cellular compartments.

The invention belongs to the technical field of subcellular level detection and analysis, and discloses a nanocapillary, a dual-circuit sensing element based on the nanocapillary and application of the dual-circuit sensing element. The nano capillary tube is of a multi-layer composite structure, and according to the double-circuit sensing element constructed based on the nano capillary tube, two independent circuits are integrated on the inner surface and the outer surface of the nano capillary tube respectively, so that accurate isolation and non-interference analysis of adjacent micro / nano-scale subcellular compartments are achieved; the design of the nanocapillary and the dual-circuit sensing element constructed based on the nanocapillary can be used as a multifunctional tool for researching the interaction between an organelle and surrounding cytoplasm.

The invention provides recombinant yarrowia lipolytica for producing beta-elemene in a subcellular compartment and a construction method and application of the recombinant yarrowia lipolytica, and belongs to the field of bioengineering. The recombinant yarrowia lipolytica is prepared by taking yarrowia lipolytica as an original strain, performing localization expression on a germacene A synthase coding gene in a liposome through a liposome localization signal Oleosin, and knocking out a peroxisome biogenic factor 10 in a yarrowia lipolytica genome, so as to obtain the recombinant yarrowia lipolytica. And inserting into a 3-hydroxy-3-methylglutaryl CoA reductase expression cassette, a farnesyl pyrophosphate synthase expression cassette, an isopentenyl pyrophosphate isomerase expression cassette, a diacylglycerol acyltransferase expression cassette and a sterol acyltransferase expression cassette to obtain the recombinant expression vector. The recombinant yarrowia lipolytica disclosed by the invention can be used for efficiently producing beta-elemene.

There is provided a method of treating an autoimmune disease comprising administering to a patient a cell composition made by transducing cells with a mixture of vectors, the mixture comprising: (i) a first vector which expresses a first chimeric antigen receptor (CAR) which binds CD19; and (ii) a second vector which expresses a second CAR which binds B cell maturation antigen (BCMA). There is also provided a method for selectively deleting anti-BCMA CAR-expressing cells and a method for restoring the plasma cell compartment in a patient who has received such a treatment.

Provided herein are methods for: (1) rapidly depleting circulating B cells and remodeling one or more B cell compartments, (2) treating B cell-related autoimmune diseases, and (3) B cell malignancies, using B cell depleting antibodies that are directed to CD19 or CD20 and one or more target antigens. Further provided herein is a CD19 / CD20 bispecific antibody with dual Fc domains that mediate enhanced effector functions and B cell depletion. HB2198 demonstrated enhanced binding to Fcγ receptors, potent effector functions, and efficient depletion of B cells in vitro and in vivo.

The present application relates to a kind of molecular chaperone, nano-carrier and its preparation method and application.The molecular chaperone is the recombinant histidine-sulphur oxygen protein coupled with single-stranded DNA, and the cysteine residue is introduced in the protein and coupled with the single-stranded DNA of amino modification.The nano-carrier is DNA tetrahedron, and the DNA tetrahedron is obtained by self-assembly based on base pairing principle from multiple DNA single strands.DNA nano-carrier is used to deliver the molecular chaperone, can realize HTrX to specific subcellular compartment, i.e.the targeted delivery of ribosome, so that the protein in ribosome exerts biological effect, reduces the negative influence of oxidative stress on protein synthesis to reverse the aging process.The present application also discloses the application of the above-mentioned molecular chaperone and nano-carrier in preparing anti-aging drugs.

The invention relates to a system and methods for enhancing access to nuclear informational molecules, such as DNA, RNA, and proteins, by analytical biomolecules, such as transposome complexes, by treating nuclei with a nuclear permeability enhancer, and to methods of using nuclear membrane, cell membrane, and external compartmentalization approaches as contiguity preserving elements.

The application discloses an oxidation-reduction sensor for monitoring H2O2 change at a plant subcell level and a preparation method and application thereof. The sensor is a fusion protein roGFP2-PRXIIB formed by fusing oxidation-reduction sensitive green fluorescent protein roGFP2 and plant endogenous peroxidase PRXIIB. The application positions the fusion protein in different cell compartments of plants and finds that the fusion protein can be used for monitoring H2O2 change between different cell compartments. Specifically, the roGFP2-PRXIIB can monitor H2O2 change in cytoplasm, nucleus, mitochondria and chloroplast of plants triggered by an inducing agent and pathogenic bacteria effect protein; the roGFP2-PRXIIB can also monitor H2O2 change in cytoplasm of plants induced by abiotic stress ABA and NaCl. In addition, the roGFP2-PRXIIB can also detect H2O2 change in pollen tubes in the process of plant growth.

The present invention describes a whole blood assay chip that enables separation of the cellular compartment from surrounding plasma, facilitating multiplexed measurement of otherwise difficult to detect soluble factors produced upon cellular stimulation. The invention is a microfluidic chip with an incubation chamber comprising an inlet, an outlet, and a fluidic barrier. Using this platform, the cellular compartment can be stimulated with various substances and levels of immunologic biomarkers could be measured. Through a proprietary immunologic algorithm described herein, this would allow for the diagnosis of an allergic response to foods, antibiotics, and / or other drugs. Additional applications of the invention besides allergy testing could include pregnancy testing, blood type, and medical applications requiring whole blood cellular stimulation readouts.

The present disclosure relates to compositions and methods for tracking and spatially localizing a cell-expressed fusion protein within the cell (with the fusion protein optionally associated with a subcellular compartment, organelle, synapse, or the like), in a manner that minimizes any disruptive impact upon the cell, at least until the detection process is initiated. Use of transcriptomics and / or barcode nucleic acid detection is employed to assess both spatial localization of intracellularly tagged fusion proteins and to establish cell-cell connectivity, e.g., in neurons across a synapse, by associating axonal identities with individual neurons at the molecular tag and transcriptome level.

Aspects of a microneedle array containing a metal-organic structure (MOF)-vaccine biocomposite and a method for using the same are disclosed herein. The microneedle array further comprises a soluble material that dissolves when inserted into the skin of a subject, thereby releasing the MOF-vaccine biocomposite. The MOF may dissolve in an acidic environment, thereby being selected to target vaccine delivery to specific cellular compartments. Methods for preparing the MOF-vaccine biocomposite and the microneedle array are also disclosed.

The invention relates to a proximity marker conjugate of lipid nanomaterial interacting protein and application of the proximity marker conjugate, and belongs to the technical field of biology. The proximity marker conjugate of the interaction protein of the lipid nanomaterial is obtained by coupling the lipid nanomaterial and biotin ligase; the lipid nanomaterial is used for marking protein interacting with the lipid nanomaterial in vivo. Functional biotin ligase is purified and then coupled to the surface of the lipid nanomaterial, in-vivo protein crown proximity labeling is achieved, and then efficient, accurate and dynamic analysis of the lipid nanomaterial in the whole in-vivo biological interaction process is achieved. The method can capture instantaneous or weak interaction, and is high in sensitivity. The method is suitable for low-abundance proteins and membrane proteins, can reserve physiological status information when being used for marking in living cells, and is not limited by cell compartment localization.

The present invention provides novel artificial nucleic acid molecules encoding at least one antigenic peptide or protein and at least one additional sequence preferably targeting the antigenic peptides or proteins to cellular compartments of interest. Further, the invention provides (pharmaceutical) compositions or vaccines and kits comprising said nucleic acid molecules. The nucleic acid molecules, (pharmaceutical) compositions or vaccines and kits are useful for treating a variety of diseases such as cancer, infectious diseases, autoimmune diseases, allergies or graft-versus host disease.

The invention relates to a system and methods for enhancing access to nuclear informational molecules, such as DNA, RNA, and proteins, by analytical biomolecules, such as transposome complexes, by treating nuclei with a nuclear permeability enhancer, and to methods of using nuclear membrane, cell membrane, and external compartmentalization approaches as contiguity preserving elements.

The invention relates to the technical field of protein engineering, in particular to a bimolecular fluorescence complementation system triggered based on interaction of two groups of nano antibodies and epitopes and application of the bimolecular fluorescence complementation system, the bimolecular fluorescence complementation system comprises a first carrier, a second carrier, a third carrier and a fourth carrier, and a protein fragment G2 in a red fluorescent protein and split green fluorescent protein system; the second carrier comprises a nano antibody NbA and an anchoring protein; and the third carrier comprises a nano antibody NbS, and another protein fragment G1 and the protein to be detected in the split green fluorescent protein system. Two groups of different nano antibody-epitopes with high specificity interact and are combined with an endocytosis and absorption experiment to realize tracing of a specific compartment of a to-be-detected protein through complementation of two split green fluorescent protein fragments. According to the invention, non-toxic visual tracking of cellular compartments, Golgi bodies and trans-Golgi body networks / early endosomes (TGN / EE) upstream of vacuoles in living cells is realized without any drug treatment.

The invention relates to the technical field of microbial fermentation, and discloses a method for improving yield of yarrowia lipolytica astaxanthin through metabolism regulation, which comprises the following steps: firstly, preparing sebacic acid-beta-sitosterol monoester as a bifunctional metabolism regulation agent for subsequently releasing a metabolism precursor in a specific cell compartment; then carrying out growth culture on yarrowia lipolytica in a fermentation culture medium with a high carbon nitrogen ratio to obtain high-density thalli; after thalli grow into a plateau phase, adding the regulating agent into the culture medium until the final working concentration is 0.1-1.0 g / L; and finally, continuously carrying out induced fermentation culture, so that the yarrowia lipolytica efficiently accumulates astaxanthin in the cells. According to the method disclosed by the invention, a specific bifunctional regulating agent is combined with a staged culture strategy, so that fixed-point release of a metabolic precursor to a storage compartment is realized, and the technical contradiction between low utilization rate of the precursor and inhibition of cell growth is solved, so that the synthesis efficiency and the volume yield of astaxanthin are remarkably improved.