2169 results about "Intracellular" patented technology

The invention relates to the field of biological medicine, in particular to a stapled peptide targeting RAS-Raf protein interaction and application of the stapled peptide, the stapled peptide is obtained in the mode that Ac-SLHDCLMKALKVR-NH2 and Ac-SLHDCLMKALKVRGL-NH2 serve as peptide chain templates, amino acid at the i site and amino acid at the (i + 4) site are replaced with S5 and cyclized, and the nitrogen terminal is further modified with stearic acid. The stapled peptide can effectively inhibit proliferation and migration of tumor cells; the serum stability is high, and meanwhile, the cell transmembrane capability is excellent, so that target protein in cells can be better targeted, and the biological function of the target protein can be exerted. Therefore, the alphaH helical conformation of the RBD of the RAF can be stabilized through the stapled peptide and a nitrogen-terminal fatty acid modification strategy, the membrane permeability and stability of the RAF are improved, the binding force of the RAF and RAS is further improved, and proliferation and migration of various tumor cells are inhibited.

The present invention relates to various molecular constructs having at least one cellular-targeting element, one protein-targeting element, and one ubiquitin ligase recruitment element, wherein these elements are conjugated in a manner that facilitates cellular uptake and targeted degradation of specific proteins within cells. The present invention further relates to multifunctional proteolysis-targeting chimera conjugates and uses thereof for the treatment of diseases or disorders such as immune and inflammatory diseases, infectious diseases, cardiovascular diseases, endocrine disorders, and various cancers that are otherwise resistant to traditional therapeutic regimens.

Provided herein are devices, methods, and systems for in situ gene sequencing of a target nucleic acid in a cell in an intact tissue. Methods of screening a candidate agent to determine whether the candidate agent modulates gene expression of a nucleic acid in a cell in an intact tissue are also provided herein.

The invention belongs to the technical field of biochemical engineering, and provides a method for increasing L-carnosine yield of escherichia coli by increasing PRPP supply, and the yield and yield of L-carnosine are remarkably increased by up-regulating expression of a prs gene, increasing PRPP content in cells and promoting synthesis of L-histidine. Experiments show that the yield of the L-carnosine in different recombinant strains is increased by 55.56%-140%, and L-histidine and beta-alanine do not need to be added in the production process.

Systems and methods are provided for stabilization of cells and retention of biomolecules therein for further analysis. Block copolymers are inserted into membranes of biological particles, cell membranes or organelles (e.g., organelle membranes) and polymerized to form an intra-biological particle (e.g., intracellular) polymer network.

The present disclosure provides compounds according to Formulae I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIII, and XIV for use as a component in a lipid nanoparticle composition for the delivery of biological and / or therapeutic agents. The present disclosure also provides novel, stable lipid nanoparticle compositions comprising one or more biological and / or therapeutic agents, as well as methods of making and delivering lipid nanoparticle compositions.

The invention relates to an engineering bacterium of high-yield ornamental blue as well as a construction method and application of the engineering bacterium, and belongs to the technical field of natural dye synthesis by a biological method. The engineering bacterium capable of highly producing the ornamental blue, disclosed by the invention, expresses icd, bpsA, glnAY405F and gd hA; acnR, yggB, glsK, aceA and ldh are knocked out from the engineering bacteria. According to the application, yggB and aceA genes are knocked out, site-directed mutagenesis (Y405F) is carried out on a glnA gene, a glsK gene is replaced by a glnAY405F gene, an acnR gene is replaced by an icd gene, an ldh gene is replaced by a gdhA gene, and a bpsA gene is integrated, so that the formation of by-products such as lactate and succinic acid in a blue-viewing synthesis process can be blocked; the flow direction of a passage from citric acid to isocitric acid to alpha-ketoglutaric acid to glutamic acid to glutamine is improved, so that the concentration of glutamine in cells is improved, and the yield of the ornamental blue is improved.

The invention belongs to the technical field of biology, and discloses a stable protective agent, a cell antigen dry sheet and a preparation method and application of the dry sheet, the stable protective agent comprises glycerin with the volume fraction being 0.4%, gelatin with the mass volume fraction being 0.1%, EDTA with the mass volume fraction being 0.1 mM, trehalose with the mass volume fraction being 0.03% and resveratrol with the mass volume fraction being 100 [mu] M. The stable protective agent is adopted for preparing the cell antigen dry sheet, and the cell antigen dry sheet is applied to CBA detection. The stable protective agent has good permeability, can penetrate through cell membranes to enter cells, provides a protection effect inside and outside the cells, has moisture retention and oxidation resistance, can prevent cell deformation and cell fixing layer shedding, and can well maintain the stability and reliability of antigen proteins, and the prepared cell antigen dry sheet can be stored for a long time, and has a good application prospect. The antigenicity of a membrane signal index is not damaged. When a positive sample is detected, obvious green fluorescence cells can be seen, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed conveniently.

The invention belongs to the field of gene engineering, and relates to a DNA binding protein based on TraC effect protein, a fusion protein with a cytosine / adenine base editing function, an editing system and nucleic acid molecules or constructs for encoding the DNA binding protein and the fusion protein. The invention also relates to complexes and kits for gene editing comprising the fusion proteins of the invention or the nucleic acid molecules encoding them. The base editing tool disclosed by the invention can realize efficient, accurate and safe base editing in cells of various eukaryotes such as animals and plants. The unique characteristics of the base editing tool enrich the types of existing base editing tools, and the base editing tool has wide application prospects.

On one hand, the invention relates to a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material which is prepared from an S, Se-CQDs solution and a Halicin solution in parts by weight as follows: 50-120 parts of the S, Se-CQDs solution, 0.1-0.3 parts of an EDC chemical reagent, 0.1-0.3 parts of an NHS chemical reagent, 125-300 parts of the Halicin solution and 12-35 parts of a NaOH solution, and on the other hand, the invention relates to a preparation method of the sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material. The characteristics that the sulfur-selenium-doped carbon dots are uniform in size and easy to enter cells are utilized, the sulfur-selenium-doped carbon dots can cooperate with the Halicin to enter the cells to destroy the flow of protons on cell membranes, so that the sterilization and antibacterial effects are achieved, the sulfur-selenium-doped carbon dots and the Halicin synergistic antibacterial material can form a hydration layer on the surface of a wound by utilizing the oxidation resistance of the sulfur-selenium-doped carbon dots, and the antibacterial effect of the wound is improved. The adhesion of bacteria is prevented, so that the antibacterial effect is enhanced.

The invention provides a membrane protein targeted degradation agent based on cell autophagy, the degradation agent has a first structure combined with membrane protein and also has a second structure capable of inducing cell autophagy, and the first structure is connected with the second structure. According to the membrane protein targeted degradation agent based on cell autophagy, the cell autophagy mechanism is utilized for membrane protein degradation for the first time, due to the fact that autophagy is an internal mechanism of various cells, universal induction can be achieved in various cells, and the autophagy does not depend on factors such as lysosome transporter protein and E3 ligase on cell membranes; the method has the characteristics of broad spectrum and higher universality; the design route of direct connection of the first structure and the second structure is simple and easy to realize.

The invention discloses a bionic nano-composite, a preparation method thereof and application of the bionic nano-composite in preparation of a product for treating atherosclerosis, and belongs to the technical field of medicine preparation. The bionic nano compound is a metal nano enzyme Ir-TiO2 coated by an M2 macrophage cell membrane. According to the invention, the surfaces of Ir-TiO2 nanoparticles are coated with M2 macrophage membranes, so that the nanocomposite with a bionic structure is prepared. After intravenous injection, the formed bionic nanocomposite can actively target endothelial cells in atherosclerotic lesions, and release Ir-TiO2 antagonism to endothelial cells in plaques and cytoplasm of inflammatory macrophages. After the Ir-TiO2 reaches the diseased region, the Ir-TiO2 can play an enzyme role to achieve an anti-inflammatory effect, and can promote inflammatory cells such as macrophages and smooth muscle cells at the diseased region to discharge cholesterol, reduce lipid deposition in the cells, reduce the generation of foam cells and promote the growth of plaques. The nanotherapy can effectively prevent the progress of atherosclerosis lesion inflammation and relieve atherosclerosis.

The invention discloses an indole-benzothiadiazole derivative as well as a preparation method and application thereof. The general structural formula of the indole-benzothiadiazole derivative provided by the invention is shown in the specification, wherein R is selected from one of methyl piperazinyl, diethylamino, azetidinyl and pyrrolidinyl. The derivative can be specifically combined and fluorescently labeled with mitochondrial RNA in cells, and has good selectivity, high fluorescence signal-to-noise ratio and excellent anti-interference capability in a complex biological system. Meanwhile, the indole-benzothiadiazole derivative provided by the invention is simple in synthetic route, easily available in raw materials, good in stability and convenient to store and use, can be specifically combined with RNA and generate strong fluorescence response, is suitable for real-time detection and imaging of in-vitro and in-vivo mitochondrial RNA, and is an efficient mitochondrial RNA micromolecular fluorescent probe.

The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.

The invention discloses application of MIGA2 as a therapeutic target in a medicine for preventing and treating Alzheimer's disease, and belongs to the technical field of biological medicine. The MIGA2 gene is used as a therapeutic target to be applied to development, screening or preparation of drugs for preventing and treating Alzheimer's disease. After an MIGA2 overexpression plasmid is adopted to transfect an Alzheimer's disease cell model, it is found that autophagy flow in cells can be promoted and accumulation of related toxic proteins can be reduced, it is proved that MIGA2 has the neuroprotective effect on the Alzheimer's disease, and a new way and exploration direction are provided for treatment of the Alzheimer's disease.

The invention relates to a method of modulating the expression of an endogenous gene in a somatic cell derived from an induced pluripotent stem cell (iPSC), comprising differentiating an engineered iPSC capable of stable or inducible expression of a CRISPR-related protein into a somatic cell prior to introduction of a guide RNA (gRNA) sequence. The invention also relates to the engineered iPSC, cells obtained by the method and uses thereof.

The invention provides an ER targeting self-assembly AIE probe with cascade ROS / RNS generation performance, and belongs to the technical field of material chemistry, the probe is composed of four units: 1) an AIE photosensitizer OTBS used for generating ROS; 2) deriving peptide FFVLK from beta amyloid protein, and promoting the self-assembly of the peptide to form a nanofiber with a beta-sheet structure; (3) an oligomerization arginine unit RRRR which is used as an NO donor; and 4) ER targeting signal peptide KDEL to endow ER with targeting ability. Based on ER targeting and a self-assembly process, the self-assembly AIE probe is subjected to ER in-situ self-assembly in cells to form nanofibers, ROS / RNS is generated in a cascading manner under illumination, and strong ICD is induced. In addition, the tumor residence time of the photosensitizer is prolonged due to the formation of the nanofibers, and the treatment effect is further improved. The invention provides a new thought for the design of a novel light-operated II-type ICD inducer.

The instant disclosure relates to lipid particles that harbor cationic lipids, the particles found to be capable of delivering associated cargoes - particularly nucleic acid cargoes when formulated as nucleic acid-lipid particles - intracellularly to skin tissue cells when administered topically to a subject. The instant disclosure provides compositions comprising such lipid particles, optionally in association with a therapeutic agent (e.g., a therapeutic mRNA and / or nucleic acid controller system), as well as methods and kits for delivering a lipid particle-associated therapeutic agent and / or for treating or preventing a disease or disorder, e.g., a skin disease or disorder, in a subject, using one or more lipid particle compositions provided herein.

The invention discloses organic nano composite hydrogel as well as a preparation method and application of the organic nano composite hydrogel, and belongs to the technical field of biological medicine delivery. The polysaccharide-based nano prodrug in the composite hydrogel can be subjected to surface protonation under the stimulation of a tumor extracellular slightly acidic environment, so that cellular uptake is promoted, low-pH / high-concentration glutathione in tumor cells is responded, intracellular aggregation and controllable release of the nano drug are realized, the retention time of the drug in the cells is prolonged, and the bioavailability of the drug is improved. The technical effect of killing tumor cells through combined chemotherapy is achieved. According to the preparation method disclosed by the invention, the release of the polysaccharide-based nano prodrug from the hydrogel and the tissue infiltration capacity are accelerated by adopting fluorinated orthoester, the infiltration of the polysaccharide-based nano prodrug to cells is increased, and then the drug-loaded compound is encapsulated by using gellan gum, so that the slow-release effect of the polysaccharide-based nano prodrug is enhanced; therefore, the efficient enrichment of the medicine at the tumor part is realized.

The present invention relates to a fusion protein that produces point mutations in cells, its preparation and use. Specifically, the fusion protein provided by the present invention contains a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity, or is formed by a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity. The present invention also relates to the coding sequence of the fusion protein, a polynucleotide sequence containing the coding sequence, a nucleic acid construct containing the polynucleotide sequence, a corresponding host cell, a method for producing point mutations in a cell, and a kit, etc. By using the present invention, it is possible to achieve site-directed mutagenesis while obtaining high mutation efficiency and a variety of mutation combinations in a specific gene region.

The invention discloses an ecDNA (ectodeoxyribonucleic acid) identification method based on scATAC-seq (scATAC-Seq) data. The ecDNA recognition method provided by the invention can be used for accurately detecting copy number amplification, breakpoint quantity and ecDNA feature quantity on the whole cell level. According to the method provided by the invention, the specific carried annular oncogene in the tumor can be identified, the distribution characteristics of the annular oncogene in the genome can be analyzed, and the overall composition of all annular DNAs in a single cell and the specific composition of the single annular DNA can be disclosed. In addition, according to the method, the existence of the circular DNA can be verified through a dyeing experiment, and the reliability of a detection result is ensured.

The invention relates to the technical field of biomedicine, discloses mutant photo-induced ion channel ChR-2 protein, and further discloses a nucleic acid construct, an expression vector, a cell, related application and a computer model. The light-sensitive channel ChR2 protein mutant obtained by the invention has stronger light current, and the light-sensitive capability of the light-sensitive channel ChR2 protein mutant is at least improved by 100 times; besides, the invention also obtains a nucleotide sequence for coding the light-sensitive channel ChR2 protein mutant, constructs a recombinant expression vector, and obtains the light-sensitive channel ChR2 protein with higher expression quantity and stronger light sensitivity, and the light-sensitive channel ChR2 protein is very suitable for expression in cells of mammals (especially human); according to the invention, the rhodopsin in different channels is systematically studied by modifying the position G224 in the helix 6 of seven transmembrane helix motifs, which proves that the mutation of the position G224 in the helix 6 in WT ChR2 accelerates the photosensitivity of the channels, and the considered position is homologous in the helix 6 of the rhodopsin in different channels.

The invention belongs to but is not limited to the technical field of biomedicine, and discloses application of golden amide alcohol ester to preparation of a medicine for treating cholestasis. The in-vitro cell level verifies that the golden amide alcohol ester has relatively strong activity of activating FXR and CAR. The golden amide alcohol ester can relieve the cholestasis symptom of a cholestasis model mouse. The golden amide alcohol ester can obviously reduce the content of total bile acid and bilirubin in cells, relieve cholestasis and relieve hepatocyte damage. The golden amide alcohol ester can regulate and control the protein level of transporters and metabolic enzymes related to bile acid and bilirubin through double excitation of FXR and CAR, relieve cholestasis and achieve the purposes of benefiting gallbladder and removing jaundice.

The application relates to an engineering bacterium for high yield of observation blue and a construction method and application thereof, and belongs to the technical field of biological synthesis of natural dyes. The engineering bacterium for high yield of observation blue expresses icd, bpsA, glnA Y405F and gdhA; the engineering bacterium knocks out acnR, yggB, glsK, aceA and ldh. By knocking out the yggB and aceA genes, performing site-directed mutation (Y405F) on the glnA gene, replacing the glsK gene with the glnA Y405F gene, replacing the acnR gene with the icd gene, replacing the ldh gene with the gdhA gene, and integrating the bpsA gene, the application can block the formation of by-products such as lactate and succinic acid in the observation blue synthesis process, improve the flow direction of the citric acid->isocitric acid->alpha-ketoglutaric acid->glutamic acid->glutamine path, and then improve the intracellular glutamine concentration, so that the yield of observation blue is improved.

The invention provides an organic molecule cytoplasm delivery carrier as well as a preparation method and application thereof, the compound for cytoplasm delivery has the characteristic of fat-soluble aggregation, and can form electrostatic adsorption with small molecule protein goods such as RNase A (13.7 KDa), Trypsin (24KDa) and Saporin (32.8 KDa), medium molecule protein goods such as HRP (40KDa) and macromolecular protein goods such as beta-Gal (430KDa), so that the compound for cytoplasm delivery can be used for preparing a lipid-soluble protein carrier. And furthermore, transmembrane transportation is formed through the strong fat solubility property, so that the protection and intracellular delivery of carried protein molecules are realized. According to the delivery mechanism, cargo molecules can quickly penetrate through cell membranes, and carried protein molecules can enter cytoplasm to exert functions without release operation. Development of the organic molecule cytoplasm delivery carrier is expected to assist potential protein therapies with low bioavailability and off-target toxicity to complete clinical transformation, which is crucial to tumor therapy, gene therapy and the like requiring precise local administration.

The present invention relates to an RNA capping enzyme comprising or consisting of an amino acid sequence having high identity with any of the sequences of SEQ ID NO: 1-28, which can be expressed in vitro, in human cells, in mouse cells or in yeast cells, and which has the activities of RNA 5 '-triphosphatase (RT), guanylate transferase (GT) and RNA guine-N7 methyltransferase (RNMT) at the same time. The invention also relates to a mutant obtained by carrying out substitution, addition and / or deletion of amino acid on any sequence in SEQ ID NO: 1-28, a nucleotide sequence for coding the capping enzyme and / or the mutant thereof, and an expression construct or a host cell containing the nucleotide sequence. The invention also relates to a composition comprising the capping enzyme as well as a method and a reaction system for capping RNA by using the capping enzyme, a mutant or a composition thereof.

The invention provides a plug-and-play Mi3 protein nanocage fusion protein with a VNP6 tag and a preparation method and application of the plug-and-play Mi3 protein nanocage fusion protein, the fusion protein comprises the VNP6 tag, a SpyCatch003 component, a Mi3 protein nanocage and a functional protein sequence, and the VNP6 tag is located at the N end of the fusion protein. The VNP6 peptide sequence or the variant thereof is utilized to induce a vesicle or vesicle-like structure beneficial to protein folding and stabilization in escherichia coli, so that the correct folding rate and activity of recombinant protein are improved while the overall intracellular molecular crowding effect is relieved. The core idea of the invention lies in that the internal structure of cells is regulated at low temperature, and the internal microenvironment of an escherichia coli expression system is optimized, so that protein molecules under high expression load can obtain more reasonable spatial distribution, and the problems of misfolding and aggregation caused by molecular crowding are reduced.

Heterologous polypeptides expressed in a recombinant eukaryotic host cell may exhibit reduced stability and / or biological activity when expressed in an intracellular form. The present disclosure provides a heterologous polypeptide comprising at least one amino acid modifications which increases its stability and / or biological activity when expressed intracellularly in a recombinant eukaryotic host cell.