16 results about "Intracellular protein" patented technology

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.

This invention relates to the field of biopharmaceutical technology, and in particular to the application of myrotoxin A in the preparation of drugs for treating prostate cancer. This invention is the first to discover the function of myrotoxin A in inducing ferroptosis in DU145 prostate cancer cells by targeting and inhibiting ACSL3 protein, and its inhibitory effect on prostate cancer proliferation. Specific embodiments of this invention reveal for the first time that myrotoxin A can precisely utilize the unique metabolic vulnerability of prostate cancer cells, significantly downregulating the expression of ACSL3 protein in DU145 cells at the protein level. This, in turn, by relieving the inhibition of ferroptosis by ACSL3 protein, specifically induces ferroptosis in prostate cancer cells, ultimately effectively inhibiting tumor cell growth and achieving the therapeutic goal of targeting prostate cancer development.

The application discloses a preparation method of fluorine-containing Fe3O4 nano-assemblies and application of the fluorine-containing Fe3O4 nano-assemblies in intracellular protein delivery, and the preparation method comprises the following steps: preparing Fe3O4 nanoparticles and small-molecule ligands containing fluorine alkyl chains respectively, and the two are combined through ligand exchange reaction to form fluorine-containing Fe3O4 nanoparticles; the fluorine-containing Fe3O4 nanoparticles can be self-assembled with proteins to form stable complexes through the hydrophobic effect and electrostatic effect of the fluorine alkyl chains. The fluorine-containing Fe3O4 assemblies provided by the application have universality as intracellular protein delivery carriers, can effectively deliver proteins with different isoelectric points and molecular weights to tumor cells, do not affect the biological activity of the cargo proteins, have small toxicity to cells, and have good biocompatibility. The protein delivery efficiency of the assemblies is further improved under the guidance of an external magnetic field, so that the killing effect of therapeutic proteins on tumor cells is enhanced, and the assemblies have application potential in the field of tumor treatment.

Method for determining cell viability in a cell culture of biological cells for the biotechnological production of proteins, in particular antibodies, characterized in that a fluorescent dye selected from 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonate (Bis-ANS) or 8-anilinonaphthalene-1-sulfonate (ANS) is used as a detection agent, and wherein the fluorescent dye is present in the cell culture at a concentration of 0.1 µM to 10 µM, the fluorescent dye selectively penetrates cells with damaged cell membranes and binds to intracellular proteins, the labeled cells are detected by excitation in the UV range and detection of the fluorescence in the green wavelength range, and the determination of cell viability is carried out using an automated imaging fluorescence microscope.

This application discloses a muscle stem cell resuscitation and protection washing solution, a quality assessment kit, and a method of use, belonging to the field of cell biology technology. In this application, vitamin E succinate and mannan peptide are used as core antioxidant components, synergistically combined with Gynostemma pentaphyllum polysaccharide, Portulaca oleracea polysaccharide, trehalose, L-proline, and magnesium sulfate heptahydrate to construct a four-in-one resuscitation and protection washing solution that provides isotonic buffering, extracellular protection, intracellular protein stabilization, antioxidant effects, and mitochondrial stabilization. This increases the survival rate of muscle stem cells after resuscitation to over 93%. The kit integrates a multi-dimensional detection system for cell resuscitation and protection washing solution, cell viability, proliferation rate, Pax7 / MyoD double-positive stemness assessment, and cryopreservation damage, achieving closed-loop management of the entire process of muscle stem cell resuscitation, protection, and quality assessment. The double-positive rate of stemness markers reaches over 90%, providing reliable quality assurance for cell bank management and clinical applications.

The application provides a 6'-cyano modified locked nucleoside, nucleotide and nucleic acid polymer, and relates to the technical field of biotechnology.The 6'-cyano modified locked nucleoside provided by the application is in R configuration or S configuration, and is further modified to obtain a nucleotide and a nucleic acid polymer.The inventor has found that the 6'-cyano modified nucleic acid polymer has better nuclease resistance than unmodified or other modified nucleic acid polymers, and reduces the interaction between the nucleic acid polymer and intracellular proteins, so that the 6'-cyano modified locked nucleoside, nucleotide and nucleic acid polymer provided by the application have significant application value in the field of nucleic acid drugs.The preparation method of the above-mentioned locked nucleoside, nucleotide and nucleic acid polymer provided by the application is efficient, stable and practical, and can be compatible with various substituted or unsubstituted bases.

The application discloses a mutant strain of Grifola frondosa for improving yields of mycelium, protein and triterpenes and application thereof, and belongs to the technical field of microorganisms. The mutant strain of Grifola frondosa has a preservation number of CGMCC No. 42006. The mutant strain is obtained by using diethyl sulfate mutagenesis technology to treat protoplasts of Grifola frondosa, screening excellent strains of Grifola frondosa, continuously culturing the strains obtained through preliminary screening, and finally obtaining the mutant strain of Grifola frondosa with obviously improved yields of mycelium dry weight, intracellular protein and triterpenes. The mutant strain has good genetic stability and is suitable for being used as a strain for industrialized production of mycelium, triterpenes and protein of Grifola frondosa, thereby providing technical support for promoting development of the Grifola frondosa industry.

A method for determining the presence of a molecule coupled to the cytoplasmic side of a cellular membrane is disclosed. The method is implemented in a microfluidic setting and is particularly suitable for determining the presence and / or the activity of a G protein or an arrestin protein in a cell.

A method for sequencing DNA wherein a sample comprising isolated cell nuclei is contacted with an antibody that forms a covalent conjugate with a first DNA adapter, and wherein the formed antibody-DNA conjugate can bind a protein of interest and ligate to an end of a dephosphorylated DNA fragment, and wherein the sample is then contacted with a second DNA adapter that coheres to the first DNA adapter of the antibody-DNA conjugate to obtain a second DNA adapter-first DNA adapter—DNA fragment product that allows for sequencing of an amplified product.

The present invention relates to a dual-targeted anticancer agent that targets both a cancer cell-specific receptor and CP2c, and a production method therefor. The dual-targeted anticancer agent exhibits excellent target specificity by binding to a cancer cell-specific target and exhibits excellent anticancer efficacy by targeting CP2c, which is an intracellular protein, and thus can be effectively used for the treatment of cancer.

The application discloses a delivery carrier with nucleic acid time sequence release function and a preparation method and application thereof, and belongs to the technical field of nucleic acid drug delivery. The nucleic acid time sequence release delivery carrier comprises, from inside to outside, an inner water phase layer, a middle oil phase layer and an outer water phase layer; the inner water phase layer contains an aqueous solution of nucleic acid molecules; the middle oil phase layer contains a hydrophobic segment of an active targeting nucleic acid delivery polymer and an amphiphilic block copolymer; and the outer water phase layer contains a hydrophilic segment of the amphiphilic block copolymer. After the delivery nucleic acid enters a cell, the nucleic acid molecules on the surface of the delivery carrier are released first, and then the nucleic acid molecules in the middle oil phase layer and the inner water phase are released in turn, so that the time sequence regulation or protein expression of the nucleic acid is realized. By using the technical scheme of the application, the nucleic acid is released in time sequence, the protein is regulated or expressed in a certain order in the cell, the problem that a traditional nucleic acid carrier cannot time sequence control the protein expression in the cell is overcome, and the delivery carrier can be used for preparing a drug for treating a neural stem cell gene disease.

This invention discloses a siRNA or shRNA construct for inhibiting RSRC1 expression and its application, relating to the field of biomedical technology. It includes a specific nucleotide sequence targeting human RSRC1 gene mRNA; this specific nucleotide sequence can specifically bind to the coding region (CDS) or 3' untranslated region (3'UTR) of the RSRC1 gene, mediating the degradation or translational repression of RSRC1 mRNA, thereby reducing the intracellular expression level of RSRC1 protein; the construct is configured to block the direct interaction between RSRC1 protein and DVL2 protein, thereby inhibiting the activation of downstream Wnt / β-catenin signaling pathways and AKT / mTOR signaling pathways. This invention, while inducing tumor cell apoptosis, inhibiting colony formation, and reducing metastatic nodules in vivo, avoids the limitations of single-target silencing in completely blocking multi-pathway cross-talk, ultimately achieving a synergistic therapeutic effect of significantly reducing chemotherapy resistance, inhibiting metastasis of advanced ovarian cancer, and prolonging patient survival.

The present disclosure relates to compositions and methods for increasing the level of Klotho in a cell or in a subject and in particular to compositions and methods for treating diseases or conditions associated with Klotho. The methods described herein involve supplementing the level of Klotho protein in a cell by administering to the cell a nucleic acid encoding the Klotho protein.

This invention relates to the field of single-cell functional analysis and microfluidic chip technology, and discloses a kit and method for in-situ sequential quantitative detection of secreted and intracellular proteins in the same single cell. The kit includes: a microcavity array chip; a secreted protein capture antibody barcode chip and an intracellular protein capture antibody barcode chip; a plastic clamp; a bioaffinity modification reagent; secreted protein standards, intracellular protein standards, secreted protein detection antibody stock solution, and intracellular protein detection antibody stock solution; a fluorescent conjugate, antibody buffer, antibody blocking solution, cell lysis buffer, and cell washing buffer. The method of this invention, through a sequential detection process of first capturing and incubating secreted proteins, followed by in-situ lysis to capture intracellular proteins, achieves high-throughput, high-sensitivity, and high-specificity in-situ sequential quantitative detection of secreted and intracellular proteins in the same single cell without cell transfer or loss of spatial location information. It can establish a correlation analysis of "intracellular signaling pathway activation - external functional output" in single cells, providing a powerful tool for disease mechanism research, drug screening, and immunotherapy evaluation.

The application provides a 6'-cyano modified locked nucleoside, nucleotide and nucleic acid polymer and a preparation method and application thereof, and relates to the technical field of biotechnology.The 6'-cyano modified locked nucleoside provided by the application is in R configuration or S configuration, and is further modified to obtain a nucleotide and a nucleic acid polymer.The inventor has found that the 6'-cyano modified nucleic acid polymer has better nuclease resistance than the unmodified or other modified nucleic acid polymer, reduces the interaction between the nucleic acid polymer and intracellular proteins, and the S configuration of the 6'-cyano modified nucleic acid polymer is more significant than the R configuration in improving the nuclease resistance of the nucleic acid polymer and reducing the interaction between the nucleic acid polymer and intracellular proteins, so that the 6'-cyano modified locked nucleoside, nucleotide and nucleic acid polymer provided by the application has significant application value in the field of nucleic acid drugs.

The present application relates to a kind of enrichment type crosslinking agent transport carrier for in vivo protein complex in situ analysis and its preparation and application in in situ protein complex analysis.In the method, according to the characteristics of enrichment type crosslinking agent, enrichment type crosslinking agent transport carrier is designed and prepared, and the enrichment type crosslinking agent transport carrier is prepared by emulsion solidification, sequential nanometer precipitation, film hydration method, and based on this, in vivo in situ analysis method for analyzing intracellular protein complex under physiological condition is established.Crosslinking agent transport carrier releases crosslinking agent in situ, extracts protein complex, and then obtains crosslinking peptide segment by denaturation, reduction, alkylation, enzymatic hydrolysis protein complex, enrichment material is used to enrich crosslinking peptide segment, and mass spectrometry technology is combined, to realize the analysis of protein complex at the level of living body.