28 results about "Genetic transfection" patented technology

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 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 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.

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.

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.

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.

An effective gene therapy technique that can transfect a gene to a cell in an organ or a tissue in a subject in a safe, effective, and durable manner is disclosed herein. The methods disclosed herein can induce expression of a nucleic acid payload in target cell(s) of a mammalian subject by administering to the subject a linear double stranded DNA (dsDNA) construct covalently closed at each of its ends by DNA loops, the linear dsDNA construct comprising the nucleic acid payload; administering to the subject a plurality of microbubbles; and administering to the subject, in proximity to the target cell(s) an effective amount of an ultrasound energy, thereby inducing the formation of pores the target cell(s) by disrupting the plurality of microbubbles to result in introduction of the dsDNA into the target cell(s) and expression of the nucleic acid payload in the target cell(s).

The invention discloses an amino acid modified poly (2-hydroxypropyleneimine) gene vector and a preparation method thereof. The method comprises the following steps: adding EDC / NHS activated amino acid into a PHP solution, adjusting the pH value of a reaction mixture to 5.5 + / -0.5, and reacting at normal temperature to prepare the amino acid modified PHP gene vector. Compared with a traditional PEI material, the amino acid modified PHP gene vector disclosed by the invention is higher in gene transfection efficiency and stronger in transfection performance.

Accordingly, the present disclosure provides a population of genetically engineered mesenchymal stem cells (MSCs), comprising an expression vector comprising an Akt or HGF gene and a PD-L1 gene. Also provided is a method for synergistically increasing survival status and immunomodulatory ability of an MSC or enhancing proliferation of an MSC, comprising transfecting an MSC with an Akt or HGF gene and a PD-L1 gene and a method for preventing, ameliorating and / or treating an ischemia condition, enhancing neuroregeneration or reducing neuronal death, comprising administering an effective amount of a population of genetically engineered MSCs of the present disclosure to a subject in need thereof.

The subject invention is directed to a mixed cell composition to generate a therapeutic protein at a target site by providing a first population of mammalian cells transfected or transduced with a gene that is sought to be expressed, and a second population of mammalian cells that have not been transfected or transduced with the gene, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.

Provided are a UV light-responsive hyperbranched poly-β-amino ester having high-efficiency gene delivery ability and a preparation method and application thereof; said poly-β-amino ester uses 4-amino-1-butanol, 2-nitro-1, M-phthaloyl 3-diacrylate, trimethylolpropane triacrylate, and 1-(3-aminopropyl)-4-methylpiperazine as raw materials, is polymerized by means of the “A2+B3+C2” Michael addition method, causing it to have a hyperbranched structure. In comparison with a linear structure, the branched structure enhances the interaction between the polymer and the nucleic acid molecule, significantly improving gene condensation ability, while also increasing cellular uptake by means of enhancing the interaction with the cell membrane. The poly-β-amino ester has a UV-responsive group on the backbone chain; under UV light irradiation, the poly-β-amino ester can be rapidly degraded after endocytosis, and releases the encapsulated genes, and achieves efficient gene transfection and reduces material toxicity. The invention has good prospects for development in the field of biomedical materials, and particularly in gene delivery.

The function of enhancing stem cells is accomplished with a specific transfected human gene type into a stem cell of choice to allow for the regenerative potential of the selected stem cell to be greatly enhanced by combining both gene and stem cell therapies together. Specific genes for selected protein production are cultured with a stem cell of choice which then produces a supernatant composed of an exosome-gene complex. This exosome-gene complex can also be used as a therapeutic gene therapy on its own or in combination with a stem cell gene complex together for a specific cellular effect, organ or tissue of regenerative effect or general systemic result.

A novel cationic lipid has a structure as represented by general formula (1) and specifically relates to a nitrogen-branched cationic lipid, and a liposome containing the cationic lipid, and a nucleic-acid pharmaceutical composition containing the liposome, a preparation method and application thereof, wherein, the definition of each symbol in the formula (1) is as defined herein. The cationic liposome containing the cationic lipid as represented by formula (1) can improve the loading rate and transport efficiency of nucleic-acid drugs. The formulation of the cationic liposome nucleic-acid pharmaceutical composition has good cell compatibility and higher gene transfection capability, and can improve the treatment and / or prevention effects of nucleic-acid drugs.

The application discloses a polymer containing a chemotherapy drug SN38 and a macrocyclic polyamine, and a preparation method and application thereof, and belongs to the technical field of polymers.The polymer containing the chemotherapy drug SN38 and the macrocyclic polyamine can release the SN38 under esterase conditions; can effectively condense siRNA to form uniform spherical nanoparticles; the nanoparticles formed by compounding with auxiliary lipids (DSPE-PEG-iRGD) have excellent tumor targeting and selectivity; under hypoxic conditions, the gene transfection capacity can be improved through a photochemical internalization effect to promote lysosome escape, and the knockdown rate of Bcl-2 protein is as high as 76%; the proliferation of HCT116 cells in the mouse body is significantly inhibited through delivery of Bcl-2 siRNA and photodynamic therapy, and the tumor inhibition rate is as high as 95%. The polymer can be used for preparing a non-viral gene carrier, as a photosensitizer, and for preparing a carrier for delivering the chemotherapy drug SN38 in tumor treatment.

The invention provides an establishment method and application of a flavidae gill filament cell line. The preservation number of the cell line is CGMCC (China General Microbiological Culture Collection Center) No.46720. The flavidae gill filament cell line established by the invention has good hereditary stability, and continuous passage of more than 100 generations is realized. A gene transfection experiment verifies the efficient transfer and expression ability of the cell line to exogenous genes, can provide abundant cell resources for the research of gene functions of the Flavonus flavipes and the related research of molecular biology and the like, and has a wide application prospect.

A targeting vector includes an extracellular vesicle in which a dopamine transporter antibody is bound to a transmembrane protein, the extracellular vesicle is secreted by a cell genetically transfected with a vector gene, and at least a portion of the vector gene includes SEQ ID No: 1. The targeting carrier provided by the invention can be used for loading a medicine to pass through a blood brain barrier, can achieve specific binding aiming at dopamine nerve cells, and achieves excellent effects of regulating secretion of marker protein of Parkinson's disease and delaying the course of Parkinson's disease.

The application discloses a cyanobacterial nanogene carrier and a preparation method thereof. By irradiating cyanobacteria under appropriate wavelength light, toxic active oxygen is generated in the cyanobacteria, lysosomes are destroyed, and the gene carrier trapped in the lysosomes is released into the cytoplasm, so that the biological function of the gene can be better exerted, and the problems of the gene carrier in the prior art, such as cationic lipids, cationic polymers, difficult lysosome escape and low gene transfection efficiency, are overcome. Meanwhile, the preparation process is complex, other substances are easily introduced, and the safety is low, and the like are overcome, so that the application is more suitable for efficient delivery of the gene carrier, an effective treatment method is provided for treatment of tumors and the like, and the treatment of cancer and the like is beneficial.

The invention provides a transfection mixture preparation device, a gene transfection system and a related method, and relates to the technical field of molecule and cell biology. The transfection mixture preparation device comprises a workbench and a bag body, the bag body is a cuboid-like whole and can be divided into a nucleic acid bag, a reagent bag and a connecting bag according to functions, the two ends of the connecting bag are communicated with the nucleic acid bag and the reagent bag respectively, the nucleic acid bag is used for containing exogenous nucleic acid diluent, and the reagent bag is used for containing transfection reagent diluent. The reagent bag can move to a position higher than the nucleic acid bag under the action of external force, so that transfection reagent diluent in the reagent bag flows into the nucleic acid bag through the connecting bag and is mixed with exogenous nucleic acid diluent, and finally a transfection mixture is obtained. Therefore, the transfection reagent diluent is added into the exogenous nucleic acid diluent in a pouring manner, and the volume of the reactor is not seen in the process, so that the transfection mixture has very short uniform mixing time all the time, and the transfection efficiency is ensured.

The invention belongs to the technical field of gene therapy drugs, and particularly relates to an LDLR gene drug and application thereof. The LDLR gene therapy composition comprises an LDLR gene transfer construct and a cationic polymer carrier, the LDLR gene transfer construct comprises: a nucleic acid encoding an LDLR protein or a functional fragment thereof; the LDLR protein has an amino acid sequence as shown in SEQ ID NO.1, or comprises an amino acid sequence which has at least 90% of identity with the amino acid sequence as shown in SEQ ID NO.1, and has a variant with the same function as the amino acid sequence as shown in SEQ ID NO.1; the cationic polymer carrier is a branched cationic carrier SS-HPT modified by vitamin A molecules. The LDLR gene drug is subjected to gene transfection in a non-virus form, and has the advantages of extremely low cytotoxicity and repeatable administration; lDLR plasmids are delivered in a targeted manner, so that the purposes of treating LDLR gene defects and preventing and treating atherosclerosis are achieved.

The invention discloses a fluorinated cationic polymer, a preparation method and application of the fluorinated cationic polymer in gene transfection. According to the method, poly (2-hydroxypropyleneimine) is modified through an ethylene oxide ring-opening reaction, poly (2-hydroxypropyleneimine) is modified through fluorocarbon epoxides with different lengths, a fluorinated poly (2-hydroxypropyleneimine) material is obtained, and the material can serve as a transport carrier for eukaryotic cell transfection. The fluorinated cationic polymer gene vector disclosed by the invention can realize high-efficiency and low-toxicity transfection of various eukaryotic cells in vitro, can implement high-efficiency and low-toxicity local administration in vivo, and has a wide application prospect.

Disclosed herein are sonoporation processes which permit for repeating sonoporation gene therapy treatments in a safe and effective manner to improve nucleic acid delivery and expression in a target cell. The methods include processes for multiplexing application of ultrasound in combination with administration of sonoactive agents and nucleic acid payloads to increase delivery of the nucleic acid payloads to a target cell in a subject, thereby increasing gene transfection and expression, and, in some cases, increasing the durability of gene expression following a sonoporation based gene therapy treatments. The methods disclosed herein can include providing ultrasound energy to multiple locations in a target tissue comprising the target cells, and which can be applied in multiple treatment sessions which can be repeated.

The application discloses an in vitro transcription system of a transformed cell of a circular theileria and a construction method thereof, and the in vitro transcription system comprises IVT-EGFP-mRNA, and the structure is 5'UTR-Cla1-Kozak-EGFP-PacI-3'UTR-polyA(120). The mRNA coding the EGFP gene is synthesized by in vitro transcription, and is subjected to capping, purification and agarose gel electrophoresis analysis. Finally, the expression of the target gene in the TaNM1 cell is verified by in vitro transfection test and indirect immunofluorescence technology. The result shows that the green fluorescent protein EGFP is successfully expressed by the in vitro transcription system coding the EGFP, which indicates that the system for transfecting the EGFP gene into the transformed cell TaNM1 of the circular theileria by the in vitro transcription method is successfully established, and the system provides important reference value for the design and research and development of the RNA vaccine of theileria annulata.