113 results about "Intracellular protein" patented technology

The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing and analyte characterization. Such polynucleotide processing may be useful for a variety of applications, including analyte characterization by polynucleotide sequencing. The compositions, methods, systems, and devices disclosed herein generally describe barcoded oligonucleotides, which can be bound to a bead, such as a gel bead, useful for characterizing one or more analytes including, for example, protein (e.g., cell surface or intracellular proteins), genomic DNA, and RNA (e.g., mRNA or CRISPR guide RNAs). Also described herein, are barcoded labelling agents and oligonucleotide molecules useful for “tagging” analytes for characterization.

Provided herein are co-assembling peptides which may form granules under stimulating conditions. Also provided herein are protein carrying granules. Further provided herein, are methods of making each of the co-assembling peptides and granules.

The invention belongs to the technical field of biological medicine, and particularly relates to an engineered cell microvesicle and a delivery system based on the engineered cell microvesicle, the system realizes efficient preparation of 1-5 [mu] m cell microvesicles, and the cell microvesicles have a large space volume and can be used for preparing the cell microvesicles. The carrier can be used for loading and delivery of target protein, polypeptide and recombinase which are specifically expressed in mother cells. The system transfects mother cells through lentivirus transfection or plasmid transfection to further produce cell microvesicles, and the microvesicles can load more goods and inherit membrane proteins of the mother cells, and can also effectively load intracellular proteins to realize effective delivery. By virtue of good structural stability, high immunogenicity and excellent biocompatibility, the cell microvesicle reduces systematic toxic and side effects of a traditional carrier, is expected to become an effective drug delivery system, and has great application potential in the field of gene therapy.

The invention provides a carrier material for supporting a protein drug. The carrier material comprises an inner core, a calcium carbonate coating layer coating the inner core and a polyethylene glycol-polyglutamic acid diblock copolymer chain segment anchored on the coating layer. The intracellular protein delivery carrier provided by the invention can effectively support various metal ions and protein drugs with the Fenton / Fenton-like reaction catalysis effect, and after the intracellular protein delivery carrier is endocytosed by cells, the protein drugs, calcium ions and the metal ions with the Fenton / Fenton-like reaction catalysis effect are released; metal ions can catalyze hydrogen peroxide in the endosome / lysosome to generate hydroxyl radicals with high oxidation activity, lipid molecules are oxidized, and a lysosome membrane is destroyed; the released calcium ions can cause mitochondrial calcium overload, improve the level of hydrogen peroxide in cells, promote generation of hydroxyl radicals in endosome / lysosome and damage of membranes of the hydroxyl radicals, induce efficient rupture of the endosome / lysosome and deliver protein drugs into cytoplasm so as to play the functions of the protein drugs.

The present invention relates to a method for producing recombinant proteins, in which a larger cellular morphology is produced by, for example, degrading tags (SsrA16 and SsrA21) by SsrA, regulating morphology-related genes (including but not limited to mreBCD, ftsABEIQWXYZ, sulA, minCDE), thereby enabling more intracellular proteins to be accommodated, thereby improving the production of recombinant proteins (such as MTG and Vip) and single cell proteins.

The invention discloses application of aloe polysaccharide in regulation and control of mitochondrial energy metabolism, and belongs to the field of new application of aloe polysaccharide. The aloe polysaccharide can regulate mitochondrial energy metabolism to be converted into anabolism, promote the content of protein and amino acid in cells and promote wound healing. The preparation method of the aloe polysaccharide comprises the following steps: removing two ends of fresh aloe barbadensis leaves, and soaking the fresh aloe barbadensis leaves with pure water for 1-2 days; washing with water and removing skin to obtain aloe gel to be ground; shearing the aloe vera gel to be ground in an automatic gel grinding device to finish gel grinding, so as to obtain aloe polysaccharide feed liquid; and putting the aloe polysaccharide feed liquid into a stirring tank, stirring at 4-15 DEG C, carrying out light treatment for at least 24 hours, and then collecting the aloe polysaccharide pervaporation membrane on the surface of the liquid in a high-temperature membrane forming tank at 75-95 DEG C, thereby obtaining the aloe polysaccharide pervaporation membrane.

The invention belongs to the technical field of microbial protein separation and extraction, and particularly relates to soluble microbial protein and a preparation method thereof. The method comprises the following steps: fermenting and culturing microbial thalli to a logarithmic phase, and centrifuging to obtain mycelia; the mycelium is freeze-dried and then smashed, and mycelium powder is obtained; the mycelium powder is placed in water to be dispersed and homogenized, and homogenized homogenate is obtained; finally, alkaline protease is added into the homogenate for enzymolysis and enzyme deactivation, and the soluble microbial protein is obtained. On one hand, microbial cell walls are directly destroyed through mechanical shearing force and collision action generated by high-speed rotation of the wall breaking machine, and intracellular proteins and active ingredients are released; on the other hand, high-pressure homogenization is combined, mechanical shear force and cavitation effect are utilized to destroy a protein compact structure and expose restriction enzyme cutting sites, and alkaline protease is combined for directional hydrolysis, so that the enzymolysis time is greatly shortened, the efficiency is improved, and the extraction rate is greatly improved.

The invention provides an organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining method, which comprises the following steps of: sealing and fixing organoid glue drops, adding a mixed solution of membrane protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin for incubation, performing membrane permeation by using a permeation agent, sealing and washing, and drying to obtain the organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining product. Adding a mixed solution of an intracellular protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin into the cell plate where the transparent organ-like glue drops are located, incubating and washing, then adding a mixed solution of a membrane protein and a fluorescent secondary antibody of the corresponding species of the intracellular protein, incubating and washing in a dark place, and finally obtaining the membrane protein / intracellular protein fluorescent antibody. And finally, adding a mixed solution of DAPI and a Duncheng phosphate buffer solution, and incubating and washing in a dark place again to obtain the membrane protein and intracellular protein immunofluorescence in-situ co-dyed organoid. According to the method, high-quality and high-fidelity membrane protein and intracellular protein in-situ co-dyeing is successfully realized, the integrity of a three-dimensional structure and the high efficiency of an experimental process are taken into consideration, and the method is convenient to operate, high in repeatability and good in universality.

Single-cell multi-omics by co-encapsulating a single cell with two beads, the first an RNA barcoding bead having barcoded mRNA capture primer oligonucleotides attached on the bead surface; and the second a DNA barcoding bead having two types of oligonucleotides releasably attached to the surface: (1) barcoded adapter oligonucleotides that are complementary to oligonucleotides bound to the transposase that are eventually incorporated into gDNA fragments and (2) polyadenylated barcoded oligonucleotides containing the same barcode sequence as the adapters. In addition, integrated analysis of RNA and protein, including intracellular protein, from individual cells using similar co-encapsulation of a single cell, an RNA barcoding bead, and with / without a specific or non-specific protein binding bead in a microwell, to avoid protein fixation by first lysing the cell to liberate intracellular contents, and then capturing protein either on a solid surface or in solution with barcoded affinity reagents.

The invention relates to the technical field of biomedicine detection, and discloses a tumor cell marker detection system for predicting the activation state of intracellular protein kinase. Comprising a marker detection module used for qualitatively detecting epithelial cell markers, mesenchymal cell markers, cell polarity markers and extracellular matrix related markers in tumor cells; the data processing module is used for performing cross validation on an epithelial cell marker, a mesenchymal cell marker, a cell polarity marker and an extracellular matrix related marker; the dynamic weighting module is used for performing dynamic weighting on different markers based on a complex system theory, and endowing epithelial cadherin with a higher weight; and the judgment module is used for determining whether epithelial intercellular substance transformation occurs or not according to the dynamically weighted marker data, and predicting the activation state of intracellular phosphoinositide 3-kinase alpha and the invasiveness of tumor cells based on the determination result of the epithelial intercellular substance transformation.

An isolated peptide for regulating intracellular protein degradation that includes a linker sequence fused to a C-end degron peptide and, optionally, a caging molecule bonded to the carboxyl group of a C -terminal alanine residue. Also disclosed is a method for temporal control of protein degradation that relies on fusing a protein of interest at its C- terminus to the degron peptide having a caging molecule, and a method for high-fidelity gene editing that utilizes an RNA-dependent endonuclease modified at the C -terminus with the caged isolated peptide. A fusion protein for high-fidelity gene editing is further provided.

Disclosed herein are screening methods for identifying cell surface receptors for progranulin. Also disclosed herein are screening methods for identifying intracellular proteins bind to progranulin.

The invention discloses a dipeptide modified polymer and application thereof in intracellular delivery of protein. The screened polymer and protein form a compound and are subjected to phase transformation, solute in a liquid phase is transformed into solid-phase particles insoluble in water, early release of the target protein is reduced, and delivery of the protein in the presence of serum is achieved. The protein intracellular delivery method provided by the invention has relatively high intracellular delivery efficiency in the presence of serum-free protein and serum, and the delivery efficiency is higher than that of a commercial protein transfection reagent PULSin; after beta-Gal and other enzyme proteins are delivered into cells, the activity can be recovered, a catalytic substrate is converted into a monitorable product, and the amount of the delivered and active recovered protein is less affected by serum protein in incubation conditions; in addition, the material has low cytotoxicity, the cell activity is higher than 90% under the experimental condition of protein delivery, and the material has good biocompatibility.

Methods and compositions for enhancing nuclear DNA delivery are provided. Aspects of the methods include contacting a cell with a DNA to be delivered to the nucleus of the cell and an agent that modulates the activity of protein Barrier-to-Autointegration Factor (BAF) in a cell. In addition, compositions, including reagents, devices and kits thereof, that find use in practicing embodiments of the subject methods are provided.

The invention provides a compound as shown in a formula (0), a pharmaceutical composition containing the compound as well as a synthesis method and application of the compound. The compound provided by the invention can obviously weaken the integrated stress response (ISR) of cells and activate the activity of eIF2B, so that protein in the cells tends to be normally synthesized, and more possible drugs are provided for diseases or diseases related to eIF2B mediated by an integrated stress response (ISR) path, and / or diseases related to regulation of the activity or level of eIF2B and the activity or level of an eIF2 pathway or an ISR pathway. # imgabs0 #

The application discloses a kind of space-time controllable PROTAC based on photo-initiated in-situ generation and application thereof.The PROTAC is divided into two components: biorthogonal reaction group and the component of target protein ligand connection, biorthogonal reaction group and the component of E3 ubiquitin ligase ligand connection, can be synthesized in situ by light control PROTAC, for targeted protein degradation.It overcomes the difficulty of PROTAC molecule from scratch synthesis, synthesis step is complicated and the like, avoids the adverse pharmacological performance and toxic side effects caused by large molecular weight in traditional PROTAC design.At the same time, only after lightening, PROTAC molecule can be generated, the target protein is degraded, tumor cells are killed, off-target toxicity is avoided, and tumor treatment effect is realized.The method for generating protein hydrolysis targeting chimera by light triggering click provided by the application can controllably and efficiently synthesize PROTAC molecules with anticancer effect, and has the prospect of precise regulation of intracellular protein expression and treatment of tumor.

The invention relates to the technical field of daily chemicals, in particular to an anti-aging composition and application thereof. The composition is prepared from a white birch bark extract, snake venom peptide and Ectoine. The white birch bark extract promotes abnormal protein removal by activating an intracellular protein ubiquitination system, and improves the skin metabolism capability; the snake venom peptide slows down expression muscle contraction through a nerve regulation mechanism, and dynamic wrinkles are effectively faded; ectoin enhances cell membrane stability, protein protection and skin stress adaptive capacity, and under the synergistic effect of Ectoin, Ectoin and skin stress adaptive capacity, the comprehensive anti-aging effects of resisting wrinkles, tightening, relieving, brightening skin color and the like can be remarkably improved.

The present application relates to the field of immunotherapy, more particularly to the field of chimeric antigen receptors (CARs). Currently, second and third generation CAR designs are quite rigid in that they combine fixed costimulatory domains in cis on the same intracellular protein domain. Trans signaling is not equivalent as costimulatory receptors have different expression levels or stoichiometry. Here, a ‘mix and match’ approach is proposed where different signaling and costimulatory domains are present on separate chains within the same CAR complex, allowing increased flexibility and control of the nature and strength of the CAR-generated signal. Also proposed are polynucleotides, vectors encoding the transmembrane polypeptide chains and cells expressing such CARs. These cells are particularly suitable for use in immunotherapy, and strategies to treat diseases such as cancer using these cells are also provided.

The subject invention provides a composition comprising an enomomer which comprises a) a carrier molecule, and b) at least one aptamer, wherein the carrier molecule is an antibody, an antigen-binding moiety, a serum protein, an intracellular protein, a messenger RNA (mRNA) or human serum albumin.

The invention discloses a non-sterilization type mildew-proof and antibacterial material capable of dynamically responding to thallus proliferation as well as a preparation method and application of the non-sterilization type mildew-proof and antibacterial material. The material comprises polyvinyl alcohol, carboxymethyl cellulose or a salt thereof and metal ions, and the polyvinyl alcohol, the carboxymethyl cellulose or the salt thereof are bridged through the metal ions; the metal ions comprise Li < + >. The material disclosed by the invention takes Na < + > released by thallus proliferation as a signal medium, releases Li < + > as required through a coordination-dissociation process, and further induces thallus cells to actively ingest Li < + > through metabolic deception of occupying a potassium ion channel by utilizing a hydration radius of Li < + > similar to K < + > according to the metabolic demand of sodium excretion and potassium ingestion in the thallus proliferation process; on one hand, bacterial cell replication lacks essential nutrient substance K < + >, and finally gradually declines; on the other hand, protein space conformation is changed and inactivated by capturing water molecules of intracellular protein, and the two aspects have a synergistic effect, so that the efficient mildew-proof and antibacterial effects of Li < + > at low concentration can be realized, and the effect is lasting.

[Problem] To provide a method for fluorescent labeling of an intracellular protein using fluorescence ON / OFF control technology which can be applied to various cells. [Solution] A method for fluorescent labeling of an intracellular protein that includes fluorescent labeling of the target protein by obtaining, in a cell, a fusion protein of a protein to be labeled and an anti-DNP (dinitrophenyl compound) antibody, bringing a compound represented by formula (I) or a salt thereof into contact with the cell, and reacting the fusion protein with the compound represented by formula (I) or salt thereof. The anti-DNP antibody in the fusion protein is an anti-DNP antibody containing a light chain containing VL-CDR1 comprising an amino acid sequence represented by SEQ ID NO: 1, VL-CDR2 comprising an amino acid sequence represented by SEQ ID NO: 2, and VL-CDR3 comprising an amino acid sequence represented by SEQ ID NO: 3 and a heavy chain containing VH-CDR1 comprising an amino acid sequence represented by SEQ ID NO: 4, VH-CDR2 comprising an amino acid sequence represented by SEQ ID NO: 5, and VH-CDR3 comprising an amino acid sequence represented by SEQ ID NO: 6, or an antigen-binding fragment thereof. The anti-DNP antibody or an antigen-binding fragment thereof is a single-chain Fv (scFv).

The invention relates to a quinoxaline derivative containing trimethyl and n-propyl as well as a preparation method and application of the quinoxaline derivative, and the derivative is 1, 6, 7-trimethyl-3-propyl quinoxaline-2 (1H)-ketone and is particularly suitable for inhibiting fusarium oxysporum. The derivative is prepared by an organic synthesis method which comprises the step of carrying out cyclization reaction on N-protected o-phenylenediamine and a carbonyl compound in the presence of trifluoroacetic acid. Experiments show that the compound has a remarkable antibacterial effect on fusarium oxysporum, and the minimum inhibitory concentration is 0.50 mg / mL. The derivatives can destroy the cell membrane structure of fusarium oxysporum and cause leakage of RNA, protein and reducing sugar in cells, so that the growth of thalli is inhibited. The invention provides a theoretical basis and an experimental basis for developing a novel efficient and low-toxicity bacteriostatic agent.

The present application relates to natural ferritin, and particularly relates to an intracellular delivery system and application thereof. The system can effectively deliver various proteins into cells and maintain the biological activity thereof, and provides important technical support for the development of protein therapy with intracellular target as core. The method exhibits significant advantages in promoting intracellular protein transport, intracellular protein interaction research and protein level regulation. The delivery system has wide applicability, is simple to operate and safe and reliable, and becomes an important tool for future intracellular protein research and application.

Provided are methods for treating cancer by administrating to an individual who has the cancer an inhibitor of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and an immune checkpoint inhibitor. Also provided are methods for promoting translation of proteins within cells such that proteins include one or more misincorporated amino acids to thereby develop new proteins that contain neoantigens. Vaccines that include the neoantigens are included. Methods for producing and detecting misincorporation of a tyrosine in proteins expressed by cells are also provided.

The present disclosure provides recombinant fusion proteins comprising an E3 ubiquitin ligase domain, a small protein binding scaffold, and an anionic polypeptide. The present disclosure also provides compositions comprising the fusion proteins disclosed herein and a cationic agent that facilitates cytoplasmic delivery of the fusion protein. The small protein binding scaffold is designed to target an intracellular protein that is targeted to be degraded intracellularly by a process mediated by the E3 ubiquitin ligase domain. The compositions disclosed herein are useful for treating cancers, inflammatory diseases, or neurodegenerative disorders.

Nucleic acid molecule libraries comprising 5' and 3' primer recognition sites and a plurality of molecules comprising a target sequence molecule and a plurality of variant sequence molecules are provided. Kits comprising the nucleic acid molecule libraries are also provided as are methods of determining protein association with a nucleotide sequence and identifying an inhibitor of protein-DNA interaction.

The present disclosure relates to a chimeric protein molecule comprising a degradation domain including an E3 ubiquitin ligase motif without lysine residues and a targeting domain comprising a substrate-binding motif which is heterologous to the E3 ubiquitin ligase motif. A linker couples the degradation domain to the targeting domain. Also disclosed are compositions as well as methods of treating a disease, substrate silencing, forming a ribonucleoprotein, screening agents for therapeutic efficacy against a disease, and methods of screening for disease biomarkers, as well as mRNA molecules, vectors, and encapsulated nucleic acid molecules encoding chimeric protein molecules.

The invention belongs to the technical field of biology, and relates to a compound for targeted degradation of OGT protein (OGT protein in tumor cells) and an application of the compound in tumor resistance. The compound is a compound as shown in a general formula (I) and a pharmaceutically acceptable hydrate or salt thereof, and the compound provided by the invention has a novel structure and has degradation activity on OGT in MCF-7 cells. The compound disclosed by the invention has an anti-tumor effect.

The invention discloses a construction method and application of a kluyveromyces marxianus mutant, and belongs to the field of synthetic biology and molecular biology. According to the method, single-stranded DNA binding protein and cytosine deaminase pmCDA1 are fused, a random base editing system is constructed, and whole-genome efficient C-T / G-A mutation is realized in kluyveromyces marxianus by taking MCM2-7 as a verification target. The system does not need a specific target or an exogenous DNA template, the mutation efficiency is high, the distribution is wide, the mutation rate is improved by 303.1 times compared with that of a control group through optimal combination, and the application potential of kluyveromyces marxianus in industrial production is expanded. The method is combined with an efficient screening system, mutant strains with the intracellular protein expression quantity remarkably improved are screened out, the constructed mutation system is not only suitable for kluyveromyces marxianus cell factory transformation, but also can be popularized to genome evolution of other yeasts or eukaryotic hosts, and a new strategy is provided for industrial microorganism breeding and protein engineering.

The invention relates to the technical field of medical compounds, in particular to a deubiquitinating enzyme targeted chimera compound as well as a pharmaceutical composition and application of the deubiquitinating enzyme targeted chimera compound. The deubiquitinating enzyme targeting chimera compound is a compound with a structure as shown in a formula (I), and / or a pharmaceutically acceptable salt thereof, and / or a racemic mixture, a hydrate, a solvate, a prodrug, an enantiomer, a diastereoisomer and a tautomer thereof, the structural formula of the formula (I) is as shown in the specification, and the definitions of X, Linker and cGAS Ligand are as shown in the specification. The compound provided by the invention increases the abundance of the cGAS protein, has an obvious effect of improving the stability of the intracellular cGAS protein, and is used for preparing medicines for treating inflammatory diseases, tumors, cardiovascular and cerebrovascular diseases and infectious diseases.