56 results about "Genetic transfection" patented technology

The invention belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody for specific recognition of type II grass carp reovirus VP4 protein, an eukaryotic expression method and application. According to the invention, a target gene S6 is transfected to an HEK293 cell for expression and purification, a New Zealand white rabbit is immunized in cooperation with a Freund's adjuvant, and the rabbit-derived recombinant monoclonal antibody capable of specifically recognizing the GCRV-II VP4 protein is obtained by using ELISA, single B cell screening and eukaryotic expression technologies. The antibody has strong specificity, provides support for further establishment of a specific II-type grass carp reovirus diagnosis technology, and has great application value in development of GCRV-II related scientific research.

The application belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody specifically recognizing type II grass carp reovirus VP4 protein, a eukaryotic expression method and application. The S6 gene in GCRV-II encodes VP4 protein, the application transfects the target gene S6 into HEK293 cells for expression and purification, cooperates with Freund's adjuvant to immunize New Zealand white rabbits, and uses ELISA, single B cell screening and eukaryotic expression technology to obtain a rabbit-derived recombinant monoclonal antibody specifically recognizing GCRV-II VP4 protein. The antibody has strong specificity, provides support for further establishment of specific type II grass carp reovirus diagnosis technology, and has great application value for development of GCRV-II related scientific research.

Therefore, this disclosure provides a genetically engineered mesenchymal stem cell (MSC) population, including an expression vector containing the Akt or HGF gene and the PD-L1 gene. It also provides a method for synergistically improving the survival status and immunomodulatory capacity of mesenchymal stem cells or enhancing their proliferation, including transfecting mesenchymal stem cells with the Akt or HGF gene and the PD-L1 gene; and a method for preventing, improving, and / or treating ischemic conditions, enhancing neurogenesis, or reducing neuronal death, including administering an effective amount of the genetically engineered mesenchymal stem cell population of this disclosure to individuals in need.

The invention discloses a method for improving transfection efficiency of chicken primordial germ cells, and belongs to the technical field of cell transfection. The invention provides application of an EGFR (epidermal growth factor receptor) activator NSC 228155 in improvement of plasmid uptake efficiency and cationic liposome transfection efficiency of chicken PGCs, and a method for efficiently transfecting the chicken PGCs is constructed. Experimental results show that compared with a control group, the gene transfection efficiency of the chicken PGCs treated by the EGFR activator NSC 228155 under a DMRIE-C transfection reagent and the gene transfection efficiency of the chicken PGCs treated by the EGFR activator NSC 228155 under a LipofectamineTM 3000 transfection reagent are both remarkably improved, which indicates that the EGFR activator remarkably improves the transfection efficiency of the chicken PGCs; and the EGFR activator does not influence the cell characteristics of the chicken primordial germ cells. The method lays a foundation for establishing an efficient gene editing chicken technical system, and has a wide application prospect.

The application relates to the technical field of biological macromolecular materials, in particular to a preparation method and application of a cationic polymer gene carrier with a fluorinated end group. The cationic polymer gene carrier with a fluorinated end group has the following structure. The fluorinated initiator is used to introduce fluorinated groups in one step to initiate the synthesis of linear polyethylene imine F-PEI with various molecular weights, the introduction of the fluorinated groups improves the gene transfection efficiency, and high-efficiency DNA transfection is realized. The gene carrier can achieve high-efficiency transfection effect in the cell transfection process, the transfection process has small toxicity to cells, gene molecules can be effectively and safely delivered into cells, and the gene carrier has the advantages of high efficiency, low toxicity, low price, simple synthesis and the like. The application has the advantages of simple reaction, easily-obtained raw materials, low preparation cost, easy control of molecular weight, high yield, simple post-treatment and suitability for large-scale production.

The invention relates to the technical field of nerve cell marking and regulation and control, in particular to a nerve cell living body selective protein modification probe capable of penetrating a blood brain barrier and application of the nerve cell living body selective protein modification probe. The protein body part specifically targets a living glutamic acid energetic neuron cell membrane, the protein body part and / or a modified part thereof can regulate and control the function of the glutamic acid energetic neuron, and a coding sequence of the protein body part comprises a sequence as shown in SEQ ID NO: 1 or a sequence having at least 85% identity with the SEQ ID NO: 1. The protein modified probe provided by the invention provides a new feasible thought for developing in-vivo nerve cell selective markers and regulation tools which are non-gene transfection of blood brain barrier penetration, are high in biological safety, are more stable and are easy to obtain and have nerve cell specific type selectivity.

The invention provides an electrode suitable for electroporation of early development chick embryos and a preparation method of the electrode. The electrode comprises an upper electrode and a lower electrode; the lower electrode comprises a root end, a middle end and a tail end which are connected with one another; the root end is a straight section which forms an angle of 35-45 degrees with the embryo; the middle end is arc-shaped, and the middle end is in a state of bending downwards and bending forwards at the same time relative to the root end; the tail end is a straight section horizontal to the embryo; the upper electrode is an electrode A suitable for small-range transfection or an electrode B suitable for large-range transfection; the electrode A comprises a straight section and a downward inclined section which are connected with each other and form an angle of 35-45 degrees with an embryo; and the electrode B comprises a straight section, a downward inclined section and a tail section which are connected with one another, wherein the straight section and the embryo form an angle of 35-45 degrees, and the tail section is horizontal to the embryo. By designing a reasonable electrode shape, accurate control of transfection area positioning is facilitated, and gene transfection in an ultra-small target range is realized.

The invention belongs to the field of biological medicine, and particularly relates to an engineered macrophage, an exosome, an engineered macrophage exosome delivery system and application thereof. The engineering macrophage is constructed by transfecting the PDCD1 gene and the Trp53 gene into the macrophage, and then the engineering macrophage is injected into the abdominal cavity of a mouse with colon cancer. The engineered macrophage continuously secretes exosomes containing expression products of the PDCD1 gene and the Trp53 gene in the abdominal cavity of a mouse and directly acts on colon cancer cells. Experiments show that the engineered macrophage has good killing ability and targeting ability, and can effectively inhibit tumor growth.

The invention discloses an in-vitro transcription system of theileria annulata transformed cells and a construction method of the in-vitro transcription system, the in-vitro transcription system comprises IVT-EGFP-mRNA, and the structure of the IVT-EGFP-mRNA is 5 'UTR-Cl-Kozak-EGFP-Pacl-3' UTR-polyA (120). According to the invention, mRNA for coding the EGFP gene is synthesized through in-vitro transcription, through capping, purification and agarose gel electrophoresis analysis, the expression condition of the target gene in TaNM1 cells is finally verified through an in-vitro transfection test and an indirect immunofluorescence technology, and the result shows that the in-vitro transcription system transfected and coded with the EGFP successfully expresses the EGFP, and the expression condition of the target gene in the TaNM1 cells is verified. The invention shows that a system for transfecting the EGFP gene into the theileria annulata transformed cell TaNM1 through an in-vitro transcription method is successfully established, and an important reference value is provided for design and research and development of theileria annulata RNA vaccines.

The present invention relates to methods for preparing carbon-based nanoparticles, also known as carbon quantum dots or carbon dots (CDs), in particular cationic carbon dots (CCDs), by incubating (1) CDs obtained from the hydrothermal treatment of an amine-free aqueous solution of citric acid with (2) polyethylenimine (PEI) (3) using stirring. The invention further relates to the use of said CCDs in the generation of gene delivery and gene transfection systems for the production of retroviral vectors.

The application relates to the technical field of new biological medical materials, in particular to a cinnamaldehyde modified high-molecular nucleic acid carrier and a preparation method and application thereof, wherein the cinnamaldehyde modified high-molecular nucleic acid carrier is a cinnamaldehyde modified polyethylene imine polymer. The high-molecular nucleic acid carrier provided by the application can significantly improve the DNA and RNA transfection efficiency in different cells, and can effectively improve the biocompatibility of the high-molecular carrier. The preparation steps of the high-molecular nucleic acid carrier are simple, the biocompatibility is good, the repeatability of batches is strong, and the high-molecular nucleic acid carrier has a wide application prospect in the development of nucleic acid carriers, gene transfection and nucleic acid treatment and the like.

The invention belongs to the technical field of biological materials, and particularly relates to an implant coating as well as a preparation method and application thereof. The invention aims to provide a material for an implant coating, which is characterized in that a cytoplasmic membrane and an oxidation response lipid peptide hybrid nanoparticle compress a gene through positive charge adsorption, and the oxidation response lipid peptide hybrid nanoparticle is a lipid peptide formed by connecting a branched basic amino acid head and a hydrophobic tail through an oxidation response bond, the hydrophobic tail part is a long-chain alkyl group or a saturated / unsaturated fatty acid chain, and the cytoplasmic membrane is an amphiphilic cytoplasmic membrane which has high functional protein expression capability after being subjected to hypoxia treatment. The invention provides an innovative combined strategy, namely fusion of gene engineering and bionic technology, and aims to improve the efficiency of coating gene transfection, actively regulate and control the peripheral microenvironment of the implant, accurately promote directional recruitment and enrichment of stem cells and effectively inhibit local inflammatory response, thereby establishing long-term stable bone-implant integration.

The application discloses an electroporation transfection protection fluid suitable for abalone embryo cell gene editing and application thereof, and belongs to the technical field of gene editing. The electroporation transfection protection fluid comprises a basic fluid and a cell protection agent; the basic fluid comprises NaCl, MgSO4, CaCl2, KCl, NaHCO3, NaBr, K2HPO4, glucose and glutathione; and the cell protection agent comprises polyethylene glycol 400. The electroporation transfection protection fluid can significantly reduce the deformity rate and mortality rate of abalone cells after electroporation technology, improve the gene transfection and gene editing efficiency, and reduce the cost of gene editing experiments, thereby laying a foundation for accelerating marine biological research and promoting environmental protection, species conservation and aquaculture.

The invention belongs to the technical field of biological gene modification, and particularly relates to transgenic moss, an ozone monitoring biosensor and a preparation method and application of the transgenic moss and the ozone monitoring biosensor. The transgenic moss is obtained by transfecting a target gene into moss cells, the target gene comprises a ZAT12 promoter, and the sequence of the gene ZAT12 is as shown in SEQ ID NO: 1. The transgenic moss is transferred into an oxygen response element ZAT12 promoter, and an ozone specific signal of the transgenic moss can be actively enhanced, so that high-sensitivity (5ppb) detection of ozone can be realized.

The invention provides a trachinotus ovatus spermatogonial stem cell line as well as establishment, cryopreservation and recovery methods and application thereof, and relates to the technical field of biologication.The method comprises the steps that tissue cells of trachinotus ovatus testis are placed in a complete culture medium to be cultured, and a fresh culture medium is replaced once every 2-4 days within 15 days before culture; culturing until the bottom of the bottle is full of cells, adding a trypsin solution to digest the cells, sucking out the digested cells every 30 seconds, respectively adding a complete culture medium for culturing for three times, screening out high-proportion spermatogonial stem cells for subsequent continuous culture, subculturing to a new culture bottle according to the proportion of 1: (1-3), and culturing to obtain the trachinotus ovatus spermatogonial stem cell line. The trachinotus ovatus spermatogonial stem cells with relatively high purity are obtained by a differential digestion method, and the trachinotus ovatus spermatogonial stem cell line is obtained by setting a passage proportion for culture and can be stably passed. And gene transfection research is carried out, so that technical support is provided for extensive research on spermatogonial stem cells of trachinotus ovatus.

The application provides a gene transfection device and a gene transfection method. The gene transfection device comprises an ultrasonic transfection chip for generating ultrasonic waves and a reaction channel in contact with the ultrasonic transfection chip to receive the ultrasonic waves, and the reaction channel is used for loading a cell solution. The device provides a novel, simple, high-throughput, fast, safe and efficient solution for cell transfection, has the characteristics of wide application range, simple operation, good repeatability, strong stability and the like, and can flexibly realize efficient transfection of adherent cells and suspension cells, and has good applicability to various difficult-to-transfect cells such as stem cells, primary cells and tumor cells.

The invention relates to the technical field of plant genetic engineering, in particular to a mutant OsPBL8 gene, a preparation method thereof and application of the mutant OsPBL8 gene in enhancing rice disease resistance. The mutant OsPBL8 gene comprises an OsPBL8 gene mutant promoter full length and an OsPBL8 gene coding sequence full length, and the nucleotide sequence of the mutant OsPBL8 gene is shown as SEQ ID No: 5. The mutant OsPBL8 gene sequence is loaded on a pZ469 (pZCB11-1300-3xMyc-GFP) vector subjected to BamHI single enzyme digestion, then agrobacterium tumefaciens is transferred, and the gene is transfected into an initial rice plant by an agrobacterium tumefaciens-mediated transformation method to form transgenic rice. By modifying the promoter of the OsPBL8 gene, the disease resistance of the rice can be remarkably improved, so that a mutant OsPBL8 gene overexpression plant still has excellent rice blast resistance in an environment with a relatively large day and night temperature difference.

The present invention provides an in situ gene transfection system based on a hydrogenated TiO2 nanotube composite coating, as well as its construction method and application. The in situ gene transfection system uses hydrogenated TiO2 nanotubes as a substrate and utilizes a layer-by-layer self-assembly technique to prepare a dopamine / graphene oxide / type IV collagen composite coating. This system can enhance the surface bioactivity of biomedical metal materials while also serving as an in situ gene transfection system for substrate-mediated local viral delivery, improving gene transfection efficiency and treatment safety, and enabling precise regulation of the treatment site. The in situ gene transfection system provides a new strategy for enhancing peri-implant epithelial sealing and preventing peri-implantitis.

The invention relates to the technical field of biological materials, in particular to lipid nanoparticles based on modified natural saponin, a preparation method and application of the lipid nanoparticles, and the modified natural saponin comprises natural saponin modified by metal complexes and / or natural saponin modified by targeting molecules. The natural saponin is selected from at least one of glycyrrhizic acid, aescin, soyasaponin, ginsenoside, ursolic acid and oleanolic acid. The modified natural saponin can effectively simulate the structural function of cholesterol compounds in lipid nanoparticles, can overcome the problems of unstable physicochemical properties of cholesterol, potential safety hazards of sources and the like, and can significantly improve the gene transfection and silencing efficiency of nucleic acid drugs as a drug carrier. And a new solution is provided for gene therapy and vaccine development. The preparation method of the lipid nanoparticles based on the modified natural saponin is mild in reaction condition, simple and convenient to operate, easy to control and beneficial to industrial production.

The present invention provides a novel cationic lipid having a structure shown in general formula (1), specifically relates to a cationic lipid containing a nitrogen heterocycle and having a polar head group derived from the nitrogen heterocycle, also relates to a liposome containing the cationic lipid, a liposome pharmaceutical composition containing the cationic lipid, its preparation and application, and the definitions of each symbol in the formula are as defined herein. A cationic liposome containing the cationic lipid shown in formula (1) according to the present invention can improve the loading rate and transport rate of drugs, especially nucleic acid drugs. A foregoing cationic liposome nucleic acid pharmaceutical composition preparation according to the present invention has good gene complexing ability and high gene transfection ability, can improve the therapeutic and / or prophylactic efficacy of nucleic acid drugs, and provides more alternative cationic lipids for the field of drug delivery. #imgabs0#

The invention discloses a chemotherapeutic drug-resistant tumor cell model as well as a construction method and application thereof. According to the construction method of the chemotherapeutic drug resistance tumor cell model, the chemotherapeutic drug resistance tumor cell model is obtained by transfecting a gene for coding a BAD mutant BADS99D to tumor cells, the BAD mutant BADS99D is a protein obtained by substituting the 99th serine Ser of a BAD protein with aspartic acid Asp, and the amino acid sequence is as shown in SEQ ID NO.1. The invention further discloses a construction method of the chemotherapeutic drug resistance tumor cell model. The phosphorylation state of a BAD gene is simulated through point mutation of a BADS99 site by a transgenic technology, so that chemotherapy drug-resistant tumor cells are constructed, and the regulation and control effects of phosphorylated BAD on the biological characteristics of drug resistance, proliferation, migration, invasion, self-renewal and the like of cancer cells are analyzed on the molecular level.

Disclosed herein are sonoporogenic methods that allow for repeating sonoporogenic gene therapy treatments in a safe and efficient manner to improve nucleic acid delivery and expression in target cells. The methods include a combination for multiplex application of ultrasound in combination with application of an acoustically active agent and a nucleic acid payload to increase delivery of the nucleic acid payload to target cells in the subject, thereby increasing gene transfection and expression, and in some cases, in some cases, the delivery of the nucleic acid payload to the target cells in the subject. A process to increase the persistence of gene expression following treatment by sonoporosis-based gene therapy. The methods disclosed herein can include providing ultrasound energy to a plurality of locations in a target tissue comprising the target cells, and can apply ultrasound energy during a plurality of repeatable treatment procedures.

The invention discloses a method and system for screening and evaluating active pharmaceutical ingredients based on in-vitro culture, and relates to the technical field of enzyme activity detection.The method comprises the steps that a cell population expressing a target metabolic enzyme is constructed through gene transfection, and a fluorescently-labeled substrate detection system is established; processing the cell population by adopting a concentration gradient to obtain dynamic response data, and analyzing an enzyme activity change rate to recognize a drug action time point; an enzyme activity recovery process is monitored after the drug is removed, a reversibility value is calculated through an activity recovery curve, and a safety value is determined by combining a ratio of a maximum tolerance concentration value to a minimum effective concentration value; and evaluating the drug activity based on the reversibility value and the safety value, and screening to obtain candidate drugs. According to the method, systematic evaluation and efficient screening of the active pharmaceutical ingredients can be realized.

Provides is engineered cells, including induced pluripotent stem cells (iPSCs) and hematopoietic cells derived therefrom, pharmaceutical compositions comprising said cells, and methods of use of same, wherein one or more genes are transfected into said cells, such that bone marrow homing is enhanced.

The invention discloses an activated release platelet drug delivery system as well as a preparation method and application thereof. The system comprises platelets and a gene transfection nano-composite entrapped in cytoplasm of the platelets, and the composite is formed by electrostatic self-assembly of reactive oxygen species (ROS) responsive cationic polymers and nucleic acid drugs. The preparation method comprises three main steps of platelet separation and purification, ROS responsive gene transfection nanocomposite preparation and platelet drug loading. The platelet is used as a natural carrier, gene drugs can be effectively entrapped and protected, fixed-point activated release of the drugs is achieved in a high-ROS microenvironment of an injured part, and excellent targeting, biocompatibility and controllable release characteristics are achieved. The system is simple and convenient in preparation process and mild in condition, is suitable for gene therapy of tissue injury diseases such as spinal cord injury, and has important clinical application value.

The invention discloses preparation and application of a hyperbranched polymer with high gene transfection capacity, and belongs to the technical field of chemical synthesis and modification. The preparation method of the hyperbranched polymer with high gene transfection capacity comprises the following steps: adding an acrylate compound and organic amine into an organic solvent, dissolving, carrying out addition reaction, cooling, adding an end-capping reagent, and continuously reacting to obtain the hyperbranched polymer. The hyperbranched polymer prepared by the invention can solve the problem that the transfection efficiency is low after the existing precursor is polymerized, the transfection capacity of nucleic acid drugs in cells is remarkably improved while the biological safety is ensured, and the hyperbranched polymer is expected to replace viral vectors to be used as mainstream nucleic acid drug delivery vectors in the market.

The utility model discloses an ultrasound mediation gene transfection device which comprises a transfection box, a supporting partition plate is fixedly connected to the lower end of an inner cavity of the transfection box, an ultrasonic generator is fixedly installed at the middle end of the bottom of the supporting partition plate, a through hole is formed in the surface of the supporting partition plate, and a transfection container is movably connected to the top of the supporting partition plate. By arranging the transfection box and the supporting partition plate, a transfection container containing cell sap can be supported in the gene transfection process, and by arranging the driving motor, the first screw rod, the fixed frame, the movable frame, the movable frame, the energy converter, the through hole, the double-shaft motor, the second screw rod, the movable plate, the push rod and the ultrasonic generator, the cell sap can be supported in the transfection process. The purpose of ultrasound-mediated gene transfection is achieved, and corresponding operation can be automatically performed on cell sap in the transfection container one by one in the gene transfection process, so that the transfection operation quality is effectively ensured.

A cell mixture for the patient-specific treatment of solid tumors comprises carrier cells with a special loading. Immunocompetent autologous cells and / or genetically transfected cells are used, loaded with a combination of immunocompetent components and / or theranostic nanoparticles. The target tissue is derived from a tumor and includes malignant tumor cells and deactivated intratumoral cells, which are subsequently separated. Liposomes are loaded with a patient- and tumor-specific combination of immunocompetent components, namely at least one stable isotope with a high affinity for (epi-)thermal neutrons and / or a β- +The process involves the use of "decay" isotopes and / or a sensitizer to perform photodynamic therapy. Additionally, the cells are loaded with theranostic nanoparticles, which also allow for monitoring of the cells and charges involved. The carrier cells are gently loaded with the loaded liposomes via passive fusion. The loading rate of the carrier cells is significantly higher than with conventional methods, and at the same time, the carrier cells are highly specific to the individual tumor of the patient.

The invention discloses a nucleic acid delivery system and a preparation method and application thereof, and belongs to the technical field of biological materials. According to the invention, a benzenesulfonamide functional group is introduced into a GSs molecular skeleton, such that a novel benzenesulfonamide functionalized gemini surfactant (NGS) is constructed. Wherein benzenesulfonamide is a specific endoplasmic reticulum targeting group, and can realize endoplasmic reticulum targeting enrichment through specific binding with a sulfonylurea receptor on an endoplasmic reticulum membrane, so that the NGS gene delivery vector is distributed towards the endoplasmic reticulum. After the NGS and the pDNA form a compound, the compound enters cells through cavette protein mediated endocytosis, then targets the endoplasmic reticulum and releases the pDNA, the pDNA is delivered into the cell nucleus by utilizing the close relation between the endoplasmic reticulum and the nuclear membrane, the gene transfection efficiency is finally improved, meanwhile, the biocompatibility is further improved by introducing the gamma-polyglutamic acid (gamma-PGA) with negative charges, and the application prospect is wide.

The invention discloses a cationic polymer, a preparation method thereof and application of the cationic polymer as a pH response charge conversion gene vector. The main chain of the cationic polymer contains a positively charged tertiary amine group and negatively charged ether bond and hydroxyl group, and the cationic polymer is synthesized by an oxa-Michael addition polymerization one-pot method of a tertiary amine alcohol monomer and a diene monomer through organic phosphazene base catalysis of hydroxyl-alkene. The cationic polymer and DNA compound is negatively charged under human physiological conditions (pH = 7.0-7.4), surface charges of the compound are converted into positive charges from negative charges under acidic conditions, and gene delivery efficiency can be improved through pH response charge conversion. A compound formed by the prepared cationic polymer and DNA has lower cytotoxicity, higher gene transfection efficiency, better serum tolerance and long circulation characteristic, and is an efficient pH response charge conversion gene delivery polymer carrier material.