40results about How to "Improve gene transfer efficiency" patented technology

The present invention provides new compositions and methods for preventing and treating pathogen infection. In particular, the present invention provides compounds having an anchoring domain that anchors the compound to the surface of a target cell, and a therapeutic domain that can act extracellularly to prevent infection of a target cell by a pathogen, such as a virus. The present invention also comprises therapeutic compositions having sialidase activity, including protein-based compounds having sialidase catalytic domains. Compounds of the invention can be used for treating or preventing pathogen infection, and for treating and reducing allergic and inflammatory responses. The invention also provides compositions and methods for enhancing transduction of target cells by recombinant viruses. Such compositions and methods can be used in gene therapy.

The present invention involves methods and compositions for increasing the susceptibility of target cells to transduction by gene transfer vectors. Specifically, it is proposed that increasing intracellular permeability in epithelial tissue increases the percentage of input vector that will transduce that target tissue. Specific examples show that receptors for retrovirus are preferentially accessible on the basolateral surface of airway epithelia, and permeabilizing such tissues results in greater infection with retrovirus. This has important implications in gene therapy, for example, to treat cystic fibrosis with the CFTR gene.

A method is disclosed for increasing the efficiency of retroviral mediated gene transfer into viable target cells, which comprises transducing the target cells by infecting the target cells with a replication defective recombinant retrovirus that infects the target cells in an aqueous medium in the presence of(a) a mixture of an effective amount of a first functional material having a retrovirus binding domain that binds said retrovirus, and an effective amount of a second functional material having a target cell binding domain that binds said target cell, or(b) an effective amount of a bifunctional material having both a retroviral binding domain which does not contain the heparin binding domain derived from human fibronectin, and a target cell binding domain, wherein the bifunctional material has a retrovirus binding domain that binds to said retrovirus and a target cell binding domain that binds to the target cell.

The present invention provides new compositions and methods for preventing and treating pathogen infection. In particular, the present invention provides compounds having an anchoring domain that anchors the compound to the surface of a target cell, and a therapeutic domain that can act extracellularly to prevent infection of a target cell by a pathogen, such as a virus. The present invention also comprises therapeutic compositions having sialidase activity, including protein-based compounds having sialidase catalytic domains. Compounds of the invention can be used for treating or preventing pathogen infection, and for treating and reducing allergic and inflammatory responses. The invention also provides compositions and methods for enhancing transduction of target cells by recombinant viruses. Such compositions and methods can be used in gene therapy.

A gene transfer efficiency enhancer of the present invention which contains a histone deacetylase inhibitor (especially compound (A)) as an active ingredient is capable of enhancing gene transfer efficiency in a gene transfer mediated by an adeno-associated virus vector while retaining the advantages of the adeno-associated virus vector.

The present invention provides a more safe and highly effective stent having functions such as anti-inflammatory action, antithrombotic action, maintenance of tissue restoration response and maintenance of endothelial regeneration. More specifically, the drug / gene eluting stent has a layer containing a gene encoding a hybrid polypeptide on the surface. The hybrid polypeptide is preferably a bound of fibronectin-derived collagen binding domain (FNCBD) polypeptide and an anti-inflammatory factor or an angiogenic factor. The uniform fine particle size capsules can directly deliver the gene encoding the hybrid polypeptide to the lesion and have benefits of reducing the given doses, and thus improving safety and efficacy and further maintaining the efficacy for a long period.

A novel growing method is provided for pluripotent stem cells such as ES cells. The method of the invention is a pluripotent stem cell growing method and gene transfer method in which pluripotent stem cells are cultured under conditions that maintain their undifferentiated state and pluripotency, the method being characterized by using a liquid medium and a culturing vessel having immobilized or coated on a substrate solid phase surface a molecule which is adhesive to the pluripotent stem cells in a fixed concentration, to grow the pluripotent stem cells in a dispersed state while maintaining their undifferentiated state and pluripotency, without using feeder cells, or to transfer and express a gene therein.

The present invention provides a kit to carry out retrovirus-mediated gene transfer into target cells. The kit contains a functional material bearing a retrovirus binding domain, another functional material bearing a target cell binding domain, an artificial substrate for incubating the retrovirus contacted with the target cells, and a target cell growth factor for pre-stimulating target cells to spur them along the cell cycle. The kit of the invention may further comprise a recombinant retroviral vector, necessary buffers, and the like.

The present invention provides a retroviral vector containing a membrane protein having a hemagglutinin activity. The present inventors constructed a retroviral vector pseudotyped by the membrane protein having a hemagglutinin activity. This viral vector showed gene transfer at a high efficiency into host cells. In particular, it was established that genes can be transferred thereby at a high efficiency into cells into which genes can hardly be transferred by the conventional techniques, for example, blood cells and hematopoietic cells including hematopoietic stem cells, and mucous cells including mucosa epithelial cells. The viral vector of the present invention is highly useful as a vector for gene therapy.

The invention relates to the fields of macromolecular substances, physical chemistry, molecular biology and virology, in particular to solution of chitosan, preparation method thereof and a method for using the same to improve the efficiency of gene transduction mediated by an adenoviral vector.

Improved methods for transferring a gene into target cells by using a retrovirus, wherein the gene transfer efficiency is improved and the target cells are efficiently transformed by binding the retrovirus to a functional substance which is immobilized on as carrier and having an activity of binding to retroviruses followed by washing; using an antibody capable of specifically recognizing cells, laminin or mannose-rich type sugar chain as a substance having an activity of binding to the target cells; pre-treating the target cells so as to inactivate transferrin receptor, or introducing a new functional group into the functional substance.

Provided is an efficiency improving agent for gene transfer to mammalian cells, a method for improving efficiency of gene transfer to mammalian cells, and a method for transforming mammalian cells. The method is characterized in that tRNA is used in combination with a lipofection reagent. Preferably, the agent may be used so that the tRNA concentration in a lipofection solution falls within the range of 3 to 50 μg / mL, and the concentration in a culture is approximately 1 / 10. More preferably, tRNA and PEG may be used in combination with a lipofection reagent. According to the present invention, gene transfer to mammalian cells with high efficiency can be achieved.

Improved methods for transferring a gene into target cells by using a retrovirus, wherein the gene transfer efficiency is improved and the target cells are efficiently transformed by binding the retrovirus to a functional substance which is immobilized on as carrier and having an activity of binding to retroviruses followed by washing; using an antibody capable of specifically recognizing cells, laminin or mannose-rich type sugar chain as a substance having an activity of binding to the target cells; pre-treating the target cells so as to inactivate transferrin receptor, or introducing a new functional group into the functional substance.

The invention relates to the preparation technology of dendrite macromolecule gene carrier, in particular, the dendrite macromolecule whose number of active groups is three times that of the old nuclear product is synthesized by designing a new nuclear molecule. The technical scheme is to use trihydroxymethylaminomethane to form a new nuclear molecule pentaerythritol-dodecylamine with pentaerythritol-quaternary acid chloride derivatives after a simple organic reaction. Compared with the products of old nuclear generations, the surface charge density and the number of active amine end groups of the new generation of polymers synthesized by the new nuclei are three times higher, which is conducive to obtaining higher gene transfer efficiency. The surface of this kind of polymer is rich in amine groups, which can combine with negatively charged DNA to form a nano-supramolecular complex, which can be used as a gene carrier to transfer plasmid DNA into cells. Cell transfection experiments show that its efficiency is equivalent to that of commonly used liposome gene transfer reagents, but its cytotoxicity is far less than that of the latter. Good application prospects.

The invention discloses a magnetic behavior nanometer cell automatic sorting device and appliance, which is characterized by the following: supplying magnetic behavior sorting system, example transferring system and computer controlling system; setting the magnetic sorting system at least with magnet sorting solution and molecule; using to separating candidate stem cell from peripheral blood or navel cord blood; adopting micro-computer and modern controlling technology; automatic-controlling for cell sorting and washing course of the whole course. This invention can be used as cell source for cytology research, medicine screening and animal experiment.

The invention relates to novel chimeric adenoviruses comprising the type 5-modified chimeric adenoviruses, in which the fiber knob domain in the adenoviruses type 5 is replaced by the adenoviruses type 35 fiber knob domain and any exogenous transcriptional regulatory regions controlling expression of the E1A and E1B genes are introduced into the region from which adenoviruses type 5 E1A transcriptional regulatory region has been removed. The chimeric adenoviruses are cytotoxic or oncolytic chimeric adenoviruses and can be utilized as, for example, pharmaceutical agents having high cytotoxic activity against intractable tumors such as malignant mesothelioma.

The invention provides a gene transfer system and method for a specific poultry primordial germ cell. The gene transfer system comprises a transposase helper plasmid containing a vasa promoter with a nucleotide sequence shown in SEQ ID NO.1. The gene transfer method comprises the steps of (1) constructing the transposase helper plasmid containing the vasa promoter or containing the vasa promoter and a universal enhancer element by using a bioengineering method; (2) constructing a transposon donor plasmid containing a target gene expression box required to be transferred by using the bioengineering method; (3) extracting the plasmids, uniformly mixing the transposon donor plasmid with the transposase plasmid according to a ratio of 1 to 2, and adjusting the final concentration of a mixed plasmid to be 500 ng-1.5 mu g / mu L; and (4) wrapping the mixed plasmid obtained in the step (3) with an embedding agent and transfecting PGCs cells in early embryo blood of poultry. Through the gene transfer system and method, efficient specific integration of exogenous genes in PGCs genomes in embryos of the poultry can be realized, so that the purpose of improving preparation efficiency of transgenetic poultry is achieved.

A method of preparing a therapeutic cell population for clinical use from a starting population of cells comprising haematopoietic stem cells, said method comprising separating a population of cells that substantially do not express CD38 but which express CD34 from the starting population of cells, and transducing the separated cell population with a vector to obtain the therapeutic cell population.

The present invention provides a gene transfer agent, a gene transfer kit, and a gene transfer method excellent in safety and transfer efficiency. Specifically, the present invention provides a gene transfer agent composition, the gene transfer agent composition including a compound represented by any one of the following formulae DL-G1 to DL-G4: DL-G1: R1R2NX(XH2)2; DL-G2: R1R2NX(X(XH2)2)2; DL-G3: R1R2NX(X(X(XH2)2)2)2; and DL-G4: R1R2NX(X(X(X(XH2)2)2)2)2 (X represents —CH2CH2CONHCH2CH2N—), in which: R1 represents an unsaturated long-chain aliphatic group; and R2 represents an unsaturated long-chain aliphatic group or a saturated long-chain aliphatic group in the formulae.

A gene transfer efficiency enhancer of the present invention which contains a histone deacetylase inhibitor (especially compound (A)) as an active ingredient is capable of enhancing gene transfer efficiency in a gene transfer mediated by an adeno-associated virus vector while retaining the advantages of the adeno-associated virus vector.

Provided is an efficiency improving agent for gene transfer to mammalian cells, a method for improving efficiency of gene transfer to mammalian cells, and a method for transforming mammalian cells. The method is characterized in that tRNA is used in combination with a lipofection reagent. Preferably, the agent may be used so that the tRNA concentration in a lipofection solution falls within the range of 3 to 50 μg / mL, and the concentration in a culture is approximately 1 / 10. More preferably, tRNA and PEG may be used in combination with a lipofection reagent. According to the present invention, gene transfer to mammalian cells with high efficiency can be achieved.

The present invention provides a gene transfer agent, a gene transfer kit, and a gene transfer method excellent in safety and transfer efficiency. Specifically, the present invention provides a gene transfer agent composition, the gene transfer agent composition including a compound represented by any one of the following formulae DL-G1 to DL-G4: DL-G1: R1R2NX(XH2)2; DL-G2: R1R2NX(X(XH2)2)2; DL-G3: R1R2NX(X(X(XH2)2)2)2; and DL-G4: R1R2NX(X(X(X(XH2)2)2)2)2 (X represents —CH2CH2CONHCH2CH2N—), in which: R1 represents an unsaturated long-chain aliphatic group; and R2 represents an unsaturated long-chain aliphatic group or a saturated long-chain aliphatic group in the formulae.

The invention discloses a synthetic method and application of water-soluble fluorescent dendrimers. Fluorescent dendrimers different in generation and carrying different functional groups are prepared from the kind of water-soluble fluorescent dendrimers by the aid of a 'divergent method' or 'convergence method'. The fluorescent dendrimers peripherally carrying ammonium salt cations have excellent water solubility and biocompatibility, are capable of entering living cells, can be combined with electronegative DNA (deoxyribonucleic acid), and can serve as genetic vectors to bring exogenous nucleic acid members DNA or RNA (ribonucleic acid) into the cells. Cellular uptake experiments show that along with increase of the generation, capabilities of penetrating cell membranes and carrying genes of the dendrimers are enhanced. The synthetic method and the application have the advantages of convenience in synthesis, high efficiency, product purification and simplicity and the like, and the synthetic dendrimers are safe, low in toxicity, excellent in water solubility and good in light stability, can serve as multifunctional cell fluorescent labeled molecules and gene vectors, and are applied to scientific researches and genetic diagnosis in the field of biological medicine.

The invention relates to a magnetic nanometer cell automatic sorting device and its application. The device is at least composed of a magnetic sorting system, a sample transmission system and a computer control system. The magnetic sorting system at least includes a magnetic sorting solution, a sorting column and an optional Moving the magnetic field, the magnetic separation solution contains gamma-Fe2O3 magnetic nanoparticles wrapped in chitosan. It can be used to separate hematopoietic stem cells from peripheral blood or umbilical cord blood, and adopts microcomputer and modern control technology to automatically control the whole process of cell sorting and washing. It can separate stem cells in batches, safely, efficiently and quickly, and obtain A large number of high-purity hematopoietic stem cells can be used as a cell source for cytological research, large-throughput drug screening, in vitro culture and animal experiments.

The present invention aims at developing a gene delivery system which has a high selectivity to a target cell and can introduce and express a gene with high efficiency, particularly developing such a system for use in a gene delivery therapy using a viral vector. The present invention provides a method for targeting a drug, which comprises the step of delivering a drug containing a therapeutic gene to a target site using an anti-PAP2a antibody.

An expanding agent for hematopoietic stem cells and / or hematopoietic progenitor cells useful as a therapy for various hematopoietic diseases and useful for improvement in the efficiency of gene transfer into hematopoietic stem cells for gene therapy is provided.A method of producing hematopoietic stem cells and / or hematopoietic progenitor cells, which comprises expanding hematopoietic stem cells by culturing hematopoietic stem cells ex vivo in the presence of a compound represented by the formula following (I), a tautomer or pharmaceutically acceptable salt of the compound or a solvate thereof (wherein R1 to R6 are as defined in the description).