48 results about "Genetic transduction" patented technology

The invention discloses attenuated, brightened and replication-controllable HSV recombinant virus, a preparation method and applications thereof. According to the present invention, thymidine kinase (TK) gene essential for replicating viruses in neurons and being a main virulence factor is knocked out by using a homologous recombination method, and subsequently a red or green fluorescent gene enhancement expression cassette is recombined into the genome of the virus to construct a series of novel recombinants viruses, wherein the toxicity is markedly low, the states of infected mice are good,the fluorescence signal is strong, and the expression of the recombinant viruses is limited at the injection site after the recombinant viruses are used in in-vivo animal center labeling; by combiningwith Cre-dependent AAV helper viruses capable of expressing TK in a compensated manner, the cell-specific transmonosynaptic loop tracing is achieved; and the recombinant HSV has wide application value in nervous system targeted gene transduction, neural network transsynaptic tracing, tumor disintegration, viral replication and pathogenesis mechanism, antiviral drug screening and other fields.

The present invention relates to chimeric biological vectors for gene trnasduction in eukaryotic cells. The invention further relates to a method for producing said chimerized vectors and a method for transduction of eukaryotic cells, in particular cells expressing integrin receptors.

Disclosed are: a gene transduction method for use in the induction of the differentiation of stem cells such as ES cells or iPS cells into hepatocytes effectively; stem cells into each of which a gene useful for the induction of the differentiation into hepatocytes is introduced; and hepatocytes produced from stem cells each having the gene introduced therein. A specific gene can be introduced into stem cells such as ES cells or iPS cells using an adenovirus vector. The effective induction of the differentiation into hepatocytes can be achieved by introducing the gene. Specifically, the effective induction of the differentiation of stem cells such as ES cells or iPS cells into hepatocytes can be achieved by introducing at least one gene selected from HEX gene, HNF4A gene, HNF6 gene and SOX17 gene into the stem cells.

The invention discloses a recombinant low-toxic herpes simplex virus system derived from a herpes simplex virustype I H129 clinical toxic strain and a construction method and application thereof. A recombinant virus constructed by a target vector is a remarkably attenuated H129 recombinant virus, the abundance of exogenous gene expression is very high, the characteristics of low toxicity and long-term high expression are shown in in vitro test or animal in vivo test, center infected animals cannot cause diseases, many neurons in brain regions are highlighted, and neuron cell bodies, axonal/dendritic fiber, dendritic spines and other fine structures are marked clearly. The low-toxic herpes simplex virus is suitable for serving as a target gene long-term and high-expression gene transductionvector and achieving long-term structure tracing of a neural circuit, and due to low toxicity, the low-toxic herpes simplex virus is also suitable for functional neural circuit analysis; in addition,the low-toxic HSV has wide application value in the aspects of nervous system targeted gene therapy, virus replication and pathogenesis analysis, animal infection model establishment, antiviral drugscreening, oncolytic therapy and the like.

This application relates to a method for converting somatic cells to Neural Stem Cells (NSCs). Moreover this application relates to a method for converting human fibroblasts, keratinocytes or adipocytes to neural stem cells based on linked steps of genes transduction and chemically defined medium induction.

The invention discloses a vivo gene transduction method and the device thereof. The device consists of a bielectrode needle provide with double syringe tubes, a high-voltage alternating current sine wave signal generator, a high-voltage direct current signal generator, and a microcomputer controlled chopping module. The invention adopts the bielectrode needle of the double syringe tubes to inject DNA into muscular tissues, and simultaneously applies excitation electric fields with specific parameters to the muscular tissues where the DNA is positioned, thereby realizing that the excitation electric field intensity of the original normal electrode device applied to the muscular tissue layer is reduced from 200-700 V/cm to 20-50 V/cm under the condition of maintaining equally transduction efficiency with the normal electrode, and greatly reducing the injury to the muscular tissues. The invention has the advantages of small size, light weight, adjustable parameters, friendly clinical application interface, etc., thereby realizing safe and efficient vivo gene transduction, and providing a new method and a new tool for further carrying on clinical gene therapy tests and researches.

An antiproliferative effect of IFN-α gene transduction in pancreatic cancer cells. The invention relates to expression of IFN-α to effectively induce growth suppression and cell death in pancreatic cancer cells, an effect which appeared to be more prominent when compared with other types of cancers and normal cells. Another aspect of the invention relates to targeting the characteristic genetic aberration, K-ras point mutation, in pancreatic cancer, and that the expression of antisense K-ras RNA significantly suppresses the growth of pancreatic cancer cells. When these two gene therapy strategies are combined, the expression of antisense K-ras RNA significantly enhanced IFN-α-induced cell death (1.3-3.5 fold), and suppressed subcutaneous growth of pancreatic cancer cells in mice. The invention also relates to a method of suppressing pancreatic cancer cells using double strand RNA formed by antisense and endogeneous K-ras RNA in combination with the anti-tumor activity of IFN-α. The invention relates to the combination of IFN-α and antisense K-ras RNA as an effective gene therapy strategy against pancreatic cancer. The invention also relates to a method of treating pancreatic cancer cells disseminated throughout the body by administering the inventive combination to localized pancreatic cancer cell tumor. The invention also relates to inducing indirect immunological antitumor activity to provide systemic immunity against pancreatic cancer cells.

A cancer vaccine containing as an active ingredient an antigen-specific dendritic cell pulsed by an HM1.24 protein, HM1.24 peptide or transduced with an HM1.24-encoding gene, or HM1.24 protein-encoding DNA or RNA.

The invention belongs to the field of tumor treatment and relates to application of a novel vesicle-like oncolytic virus in preparation of antitumor drugs. The virus Ad5sPVRCD137L infects cells capable of expressing specific membrane protein VSV-G, then a centrifugal extrusion/shearing method is utilized to enable the cells to pass through membrane holes so as to form vesicles, the oncolytic virus wrapped by the vesicles is manufactured, and the membrane protein VSV-G on the surfaces of the cells is reserved on the outer surfaces of the vesicles in the preparation process. The prepared novel vesicle-like oncolytic virus can achieve heavy targeting through the membrane protein VSV-G, and therefore, the gene transduction efficiency of the virus is improved, the neutralizing effect of an antiviral antibody can be avoided, and the yield of the virus can be improved; and a soluble fusion protein sPVRCD137L expressed by the virus can achieve the dual functions of blocking an immune detection point and activating immune costimulation. The novel vesicle-like oncolytic virus can significantly activate anti-tumor immunity and finally significantly prolong survival of mice.

The invention relates to an adeno-associated virus based and lipid ultrasound microbubble mediated novel preparation capable of obviously improving gene transfection efficiency on cells with compact structure. A structure of the novel preparation is shown as a figure 1, and consists of three parts: an adeno-associated virus, perfluoropropane, and a lipid bilayer. According to the invention, the advantage of efficient gene transfection of the adeno-associated virus vector is utilized; under the effect of ultrasound, the ''cavitation effect'' generated by the ultrasound microbubbles acts on cell membranes of the cells with compact structure to generate ''overflow pores'' and increase permeability of the cell membranes, so that adeno-associated viruses released after destruction of the microbubbles can enter blood vessel walls and even tissue spaces, thereby promoting transmembrane gene transfection. The novel preparation provided by the invention is simple for preparation and convenient for usage, and does not require special production conditions.

The application discloses a recombinant lentivirus vector for treating beta-globulin afunction, a preparation method and application. A gene element of the recombinant lentivirus vector disclosed by the application is composed of a left side lentivirus long terminal repeat, a lentivirus counter-response element, a center poly-purine sequence, a gene locus regulating and controlling sequence of a beta-globulin gene, a beta-globulin gene, of which 87-th threonine is mutated into glutamine, and a right side lentivirus long terminal repeat which are connected in order. According to the recombinant lentivirus vector disclosed by the application, through optimizing and improving the gene element, the gene transduction efficiency is increased, and the beta-globulin gene can be efficiently expressed during erythroid differentiation of hemopoietic stem cells, so that the consumption of viruses can be effectively lowered, and the treatment cost is reduced while the safety is improved. The recombinant lentivirus vector disclosed by the application provides a novel treatment tool and scheme for beta-thalassemia and sickle cell anemia caused by the beta-globulin afunction.

The invention relates to application of miR-31-5p in acute myelogenous leukemia, in particular to application of a product for detecting the miR-31-5p in preparation of a reagent, a chip or a kit for auxiliary diagnosis of subjects suffering from acute myelogenous leukemia (AML) and/or prognosis of the lifetime of the subjects. The invention also relates to an application of the miR-31 initial/precursor miRNA (prime/pre-miR-31) and/or the miR-31 mature miRNA (miR-31-5p) in preparation of a medicine for treating AML (acute myeloid leukemia). The miR-31-5p is introduced into cells through a gene transduction method, so bone marrow AML cells and leukemia stem cells (AML-LSC) can be induced to die, and the cytotoxicity of the chemotherapeutic drug cytarabine is enhanced; by expressing the miR-31-5p, the growth of AML cells and AML-LSC cells can be inhibited, and the life of animals can be prolonged. The disclosure provides a new method for diagnosis and prognosis evaluation of AML, and also provides a gene therapy drug for preparing AML.

The invention discloses a method for screening and constructing primary hepatocyte klotho gene transduction stem cells in the technical field of gene transduction stem cell screening and constructing methods. Drugs are screened by taking a T allelic site of rs648202 as a drug design target on the basis of a polymorphic DNA sequence of a klotho gene exon 4 region obtained by PCR (Polymerase Chain Reaction) amplification; and active molecules capable of adjusting the expression level of a klotho gene are screened out, polymorphic typing DNA fragment transduction is adopted based on human hepatocytes, and vector transduction is performed by sequentially adopting hTERT and pLXSN dual vectors, so that transduction of a pLXSN-hTERT expression vector for efficiently expressing hTERT is achieved, and the effect of constructing stem cells for efficiently expressing the klotho gene is achieved. In a low-solubility hygromycin double-screening culture system, the survival ability of the mesenchymal stem cells for tranducing the exogenous klotho gene is improved.