4499 results about "Genetic engineering" patented technology

This invention relates to humanized antibodies and antibody preparations produced from transgenic non-human animals. The non-human animals are genetically engineered to contain one or more humanized immunoglobulin loci which are capable of undergoing gene rearrangement and gene conversion in the transgenic non-human animals to produce diversified humanized immunoglobulins. The present invention further relates to novel sequences, recombination vectors and transgenic vectors useful for making these transgenic animals. The humanized antibodies of the present invention have minimal immunogenicity to humans and are appropriate for use in the therapeutic treatment of human subjects.

This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to a novel inducible gene expression system and methods of modulating gene expression in a host cell for applications such as gene therapy, large-scale production of proteins and antibodies, cell-based high throughput screetng assays, functional genomics and regulation of traits in transgenic plants and animals.

This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to novel substitution mutant receptors and their use in a nuclear receptor-based inducible gene expression system and methods of modulating the expression of a gene in a host cell for applications such as gene therapy, large scale production of proteins and antibodies, cell-based high throughput screening assays, functional genomics and regulation of traits in transgenic organisms.

This invention relates to the fields of genetic engineering, virus replication and gene transfer. More specifically, this invention relates to polynucleotide construct, recombinant virus, transposon, and their vectors, wherein an ori derived from a DNA virus capable of replicating in vertebrate cells is inserted into the retrovirus, allowing the retrovirus following the reverse transcription to efficiently replicate as extrachromosomal or episomal DNA without the necessity of integration into the host cell chromosome. Additionally, this invention relates to polynucleotide construct, recombinant virus, transposon, and their vectors replicating episomally without aid of an ori and related elements. Also, this invention encompasses preventive, therapeutic, and diagnostic applications employing said constructs, viruses and vectors.

The present invention provides polymers made by genetically engineering microorganisms for making a self-retaining suture. In an embodiment of the present invention the genetically engineering microorganisms synthesize polyhydroxyalkanoate (PHA) polymers. In an alternate embodiment of the invention, the genetically engineering microorganisms synthesize polybetahydroxybutyrate (PHB) polymers. In an alternative embodiment of the invention, the self-retaining sutures can be made from a copolymer such as polyhydroxybutyratevalerate (PHBV), where the genetically engineering microorganisms produces PHA polymers as the monofilament base material and a different genetically engineering microorganisms produces polyhydroxybutyratevalerate (PHBV) polymers. In various embodiments of the invention, recombinant expressed self-retaining suture materials have a melting point in the range from between approximately 40° C. to approximately 180° C. In various embodiments of the invention, recombinant expressed self-retaining suture materials have extension-to-break strength of between approximately 8% and approximately 42%.

Novel chimeric constructions and methods for their use are provided for expression of exogenous genes in a plant chloroplast. Particularly, expression is achieved by the use of a chloroplast or bacterial 5′ untranslated region in the expression cassette. The expression cassette may be integrated into the chloroplast genome by the use of chloroplast DNA flanking sequences, or may replicate autonomously if provided with a chloroplast origin of replication. Plants and cells containing the transformed chloroplasts are also provided. The constructs may be used with both monocotyledenous and dicotyledenous chloroplasts.

The present invention relates to genetically engineered attenuated viruses and methods for their production. In particular, the present invention relates to engineering live attenuated viruses which contain a modified NS gene segment. Recombinant DNA techniques can be utilized to engineer site specific mutations into one or more noncoding regions of the viral genome which result in the down-regulation of one or more viral genes. Alternatively, recombinant DNA techniques can be used to engineer a mutation, including but not limited to an insertion, deletion, or substitution of an amino acid residue(s) or an epitope(s) into a coding region of the viral genome so that altered or chimeric viral proteins are expressed by the engineered virus.

The invention belongs to the field of gene engineering and particularly relates to a method for specifically knocking out a human CCR5 (Chemokine Receptor 5) gene by CRISPR (clustered regularly interspaced short palindromic repeat-associated)-Cas 9 and SgRNA (single guide RNA) for specifically targeting the CCR5 gene as well as an intermediate carrier and application thereof. The sgRNA for specifically targeting the human CCR5 gene, prepared by the method disclosed by the invention, can be used for accurately targeting the human CCR5 gene and realizing gene knockout. The preparation method is simple in steps and good in sgRNA targeting property; the knockout efficiency of a CRISPR-Cas 9 system is high.

The invention belongs to the field of genetic engineering, and particularly relates to a method for specifically knocking out hepatitis b virus cccDNA by using CRISPR-Cas9 and sgRNA for specifically targeting the hepatitis b virus cccDNA. The invention provides a method for specifically knocking out hepatitis b virus cccDNA by using CRISPR-Cas9 and sgRNA for specifically targeting the hepatitis b virus cccDNA. The sgRNA of specific targeted hepatitis b virus cccDNA prepared according to the invention can precisely target hepatitis b virus cccDNA and realize gene knockout. A preparation method is simple in steps and good in sgRNA targeting, and the knockout efficiency of a CRISPR-Cas9 system is high.

The invention belongs to the field of genetic engineering, particularly relates to a method for human CTLA4 gene specific knockout through CRISPR-Cas9 (clustered regularly interspaced short palindromic repeat) and sgRNA(single guide RNA)for specially targeting a CTLA4 gene, and provides a method for human CTLA4 gene specific knockout through CRISPR-Cas9 and sgRNA(single guide RNA)for specially targeting a CTLA4 gene. The sgRNA(single guide RNA)for specially targeting a human CTLA4 gene, prepared through adopting the method provided by the invention, can be used for accurately targeting the human CTLA4 gene and realizes gene knockout; the preparing method has simple steps, the targeting performance of the sgRNA is good, and the knockout efficiency of a CRISPR-Cas9 system is high.

The invention relates to a gene deletion attenuated African swine fever virus as a vaccine and the vaccine, and a construction method thereof. An African swine fever Chinese epidemic strain Pig/CN/HLJ/2018 is adopted, a virulence gene of the African swine fever virus is deleted by a genetic engineering technology, and the gene deletion virus of MGF360-505R deletion and joint deletion of CD2V and MGF360-505R is obtained. Experiments show that the two virus strains can provide 100% immune protection against the African swine fever Chinese epidemic virulent strains, can be used as vaccines for safe and effective prevention and control of African swine fever in China, and have great social value.

Novel compositions and methods useful for genetic engineering of plant cells to provide expression in the plastids of a plant or plant cell of cellulose degrading enzymes.

In biology and biotechnology, electroporation is an important technique for introducing entities (DNA, RNAi, peptides, proteins, antibodies, genes, small molecules, nanoparticles, etc.) into cells. Applications range widely from genetic engineering to regenerative medicine to drug delivery. It has been demonstrated that the electrical currents flowing through cells can be used to monitor and control the process of electroporation for biological and artificial cells. In this application, a device and system are disclosed which allow precise monitoring and controlling electroporation of cells and cell layers, with examples shown using adherent cells grown on porous membranes.

The invention discloses a method for amplifying NK cells of human beings under the condition of in vitro culture, which is characterized in that a peripheral blood mononuclear cell (PBMC) is taken asthe original culturing material, and the PBMC is cultured together with an engineering cell which is built by adopting the genetic engineering method and is used for stimulating the growth of the NK cell. The built engineering cell which is used for stimulating the growth of the NK cell expresses several cytokines (IL-2, IL-12, IL-15, IL-18, 4-1BB) which can promote the growth of the NK cell on the surface of the K562 cell; after irradiation and inactivation with gamma-rays, the engineering cell is cultured with the PBMC in vitro, as a result, the amplification effect of the stimulation methodin the invention is hundreds of times stronger than that of the currently universal method in which soluble cytokines are added purely to the culture solution; and after cultured for 3 weeks, the non-NK cells in the PBMC are mainly dead and disappeared, the NK cells are proliferated in great quantity, the purity of the NK cells reaches over 96% and the total number of the NK cells is amplified byover 1500 times.

The present invention relates to the field of biotechnology or genetic engineering. More specifically, the present invention relates to a multiple inducible gene regulation system that functions within cells to simultaneously control the quantitative expression of multiple genes.

Normal cells, such as fibroblasts or other tissue or organ cell types, are genetically engineered to express biologically active, therapeutic agents, such as proteins that are normally produced in small amounts, for example, MIS, or other members of the TGF-beta family Herceptin(TM), interferons, andanti-angiogenic factors. These cells are seeded into a matrix for implantation into the patient to be treated. Cells may also be engineered to include a lethal gene, so that implanted cells can be destroyed once treatment is completed. Cells can be implanted in a variety of different matrices. In a preferred embodiment, these matrices are implantable and biodegradable over a period of time equal to or less than the expected period of treatment, when cells engraft to form a functional tissue producing the desired biologically active agent. Implantation may be ectopic or in some cases orthotopic. Representative cell types include tissue specific cells, progenitor cells, and stem cells. Matrices can be formed of synthetic or natural materials, by chemical coupling at the time of implantation, using standard techniques for formation of fibrous matrices from polymeric fibers, and using micromachining or microfabrication techniques. These devices and strategies are used as delivery systems via standard or minimally invasive implantation techniques for any number of parenterally deliverable recombinant proteins, particularly those that are difficult to produce in large amounts and/or active forms using conventional methods of purification, for the treatment of a variety of conditions that produce abnormal growth, including treatment of malignant and benign neoplasias, vascular malformations (hemangiomas), inflammatory conditions, keloid formation, abdominal or plural adhesions, endometriosis, congenital or endocrine abnormalities, and other conditions that can produce abnormal growth such as infection. Efficacy of treatment with the therapeutic biologicals is detected by determining specific criteria, for example, cessation of cell proliferation, regression of abnormal tissue, or cell death, or expression of genes or proteins reflecting the above.

The invention relates to the technical field of genetic engineering, in particular to a method for specifically knocking out a human PCSK9 gene by virtue of CRISPR-Cas9 and sgRNA for specifically targeting the PCSK9 gene; through high-throughput cloud computing and designing, a group of sgRNA for specifically knocking out the specifically targeted PCSK9 gene in the human PCSK9 gene by virtue of the CRISPR-Cas9 is synthesized; meanwhile, each sgRNA is linked to a linear pCG-U6-SgRNA plasmid, so that a carrier is obtained, and a pair of forward and reverse sgRNA oligonucleotide carriers, together with the pCG-NLS-FLAG-Cas9 plasmid, are used for successfully transfecting cells, so that the PCSK9 gene is knocked out; the method has the advantage that the large-scale production of the sgRNA can be achieved just by synthesizing a few of polynucleotide segments; and the preparation method is simple and easy to implement, good in sgRNA targeting properties and high in knocking-out efficiency on the CRISPR-Cas9 system.

The invention discloses a water-replenishing repairing cosmetic as well as a preparation method and application thereof. In the invention, a skin water-replenishing repairing cosmetic with an excellent effect is prepared by combining traditional cosmetic effective raw materials with genetic engineering product biotic factors and adopting a vacuum freeze-drying technique. The cosmetic disclosed bythe invention comprises water-replenishing repairing freeze-dried powder and water-replenishing repairing base liquid, wherein the freeze-dried powder comprises the following main components: grape seed extract, hydrolyzed red alga extract, low-molecular-weight hyaluronic acid, hydrolyzed collagen, recombinant human keratinocyte growth factor, recombinant human epidermal cell growth factor and micromolecular heparin sodium; and the base liquid comprises the following main components: hyaluronic acid (HA) with molecular weight of 1500000-2500000, methyl propanediol, honey extract, PCA (pyrrolidone carboxylic acid) sodium, dissolved proteinase, 1,2-hexylene glycol, oat beta-glucan, soybean isoflavone and the like. The water-replenishing repairing cosmetic disclosed by the invention can be used for daily skin nursing, can also be used through introduction by virtue of ultrasonic and other methods, and has the beautifying effects of replenishing skin water, repairing damaged cells caused by water deficiency and enhancing skin elasticity.

The invention belongs to the technical field of animal virology and genetic engineering, relating to pseudorabies virus which lacks parts of virulence gene of min strain A of pseudorabies virus, and the process for preparing vaccine with the virus. The DNA recombinant technology is employed to artificially remove all or parts of albumen code area of a plurality of genes of TK, gE, gI, 11K, and 28k which are unnecessary genes and duplicate virus of viruses and influence the virulence in the pseudorabies virus, thereby lowering the virulence of the pseudorabies virus, achieving attenuated vaccine strains SA215 of which the attenuation genetic engineering is lack, preparing vaccine with SA215 virus, and immunizing animals to effectively prevent and control the occurrence and prevalence of the pseudorabies.

The invention relates to storable medicaments produced from pharmaceutical active ingredient preparations which are virus safe. Said medicaments contain at least one intact therapeutic protein obtained from plasma or by means of genetic engineering, as an active pharmaceutical substance. Said active ingredient preparations contain active enzymes, especially proteases, which are either free or bound to the substrates thereof and act against the therapeutic protein(s) present.

The invention provides non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing transgenic cells and mammals. Certain aspects of the invention include chimeric antibodies, humanized antibodies, pharmaceutical compositions and kits. Certain aspects of the invention also relate to diagnostic and treatment methods using the antibodies of the invention.

This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to a novel ecdysone receptor/chimeric retinoid X receptor-based inducible gene expression system and methods of modulating gene expression in a host cell for applications such as gene therapy, large-scale production of proteins and antibodies, cell-based high throughput screening assays, functional genomics and regulation of traits in transgenic organisms.

This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to a novel ecdysone receptor/invertebrate retinoid X receptor-based inducible gene expression system and methods of modulating gene expression in a host cell for applications such as gene therapy, large-scale production of proteins and antibodies, cell-based high throughput screening assays, functional genomics and regulation of traits in transgenic organisms.

The invention relates to a cell line gene knock-out method for obtaining large fragment deletion through a CRISPR/Cas9 system, and belongs to the field of genetic engineering and genetic modification. A pX458 vector is modified, so that the pX458 vector is provided with DsRed2 and ECFP (Enhanced Cyan Fluorescence Protein); then, a plurality of specific sgRNA sites are designed by aiming at a target gene and are connected into the modified vector; after the cell line transfection, cell groups are sorted by a flow cytometry; single cells subjected to genome editing can be very fast obtained; then, a single-cell DNA (Deoxyribonucleic Acid) sequence is subjected to PCR (Polymerase Chain Reaction) amplification; and single cells with large fragment deletion can be picked out from the single cells through gel electrophoresis. The CRISPR/Cas9 system, the flow cytometry single-cell sorting and fluorescent protein screening on the expression vector are combined, so that the positive monoclonal cells with the large fragment deletion can be obtained in a short time; and the work efficiency of the cell line gene knock-out is greatly improved.

Disclosed are methods for producing fusion proteins with the heterodimeric cytokine, interleukin-12. In order to insure that the proper ratio of fused and non-fused subunits are obtained in the fusion protein, a specific stepwise approach to genetic engineering is used. This consists of first expressing the non-fused p40 IL-12 subunit in a production cell line, followed by or simultaneously expressing in the same cell, a second recombinant fusion protein consisting of the fused polypeptide linked by a peptide bond to the p35 subunit of IL-12. Molecules containing the p35 fusion protein cannot be secreted from the transfected mammalian cell without first complexing in a one to one ratio with the p40 subunit, thus ensuring the production of active heterodimeric fusion proteins.

A fungal microorganism can be engineered by means of genetic engineering to utilise L-arabinose. The genes of the L-arabinose pathway, which were unknown, i.e. L-arabinitol 4-dehydrogenase and L-xylulose reductase, were identified. These genes, together with the known genes of the L-arabinose pathway, form a functional pathway. This pathway can be introduced to a fungus, which is completely or partially lacking this pathway.

The invention relates to the technical field of genetic engineering, and particularly relates to a guide RNA (gRNA) sequence based on a CRISPR/Cas9 system and combinations thereof, as well as gRNA for a specific targeted knockout hepatitis B virus cccDNA P gene. In the invention, 30 gRNAs are designed according to design principles of CRISPR/Cas9, have a sequence table as shown in SEQ ID NO.1-30, and are constructed on a PX458 vector, to screen out four more efficient gRNAs. The CRISPR/Cas9 system guided by the four gRNAs and combinations thereof is utilized in a human hepatocellular carcinoma cell line (HepG2.2.15) to effectively knock out the human hepatitis B virus cccDNA P gene. The gRNA of the specific targeted hepatitis B virus cccDNA prepared in the invention can accurately target the hepatitis B virus cccDNA and realize gene knockout. The preparation method is simple in operation; the gRNA has a good targeting property; the CRISPR/Cas9 system is high in knockout efficiency.

The invention relates to an anti-human transferrin receptor human antibody and the application, which pertains to the field of molecular biology. The anti-human transferrin receptor human antibody has a single-chain antibody (scFv) amino acid sequence which is shown in a sequence table SEQ ID No.1. The anti-human transferrin receptor human antibody and the application have the advantages that: the human antibody solves the problem of immunogenicity of the target antibody, thus allowing the human antibody to carry out the long-term repeated drug administrations in the human body; the method for screening a genetic engineering antibody in a large-capacity antibody library which is adopted by the invention is easier than the monoclonal antibody preparation, the large-scale production is easy, the molecule of the antibody is small, and the antibody can enter tissues deeply, thus having obvious advantages. A pET-22b (plus) vector which is adopted by the invention contains a stronger T7 promoter, which can ensure the higher expression level of the target antibody. The induced expression conditions after the optimization can ensure that the antibody is expressed in a soluble form and the activity of protein is higher. The 3' end of the pET-22b (plus) vector is provided with six His(histidine) labels, which can easily use the Ni-NTA agarose to carry out affinity chromatography and purification.

A high flux of metabolites from pyruvate to 2,3-butanediol in Lactobacillus plantarum was achieved through genetic engineering. Substantial elimination of lactate dehydrogenase activity in the presence of heterologously expressed butanediol dehydrogenase activity led to 2,3 butanediol production that was at least 49% of the total of major pyruvate-derived products.

The invention provides a technological flow with versatility and high efficiency for reforming human serum albumins and fusion proteins of human serum albumins by separating and purifying target proteins. The steps of the method are as follows: the inorganic salt culture medium is used to acquire the genetic engineering microzyme fermented supernatant containing target proteins, and the serum-free culture medium is used to acquire a culture solution of reforming vertebrate cells, and ion exchange type affinity column chromatography medium Capto MMC fillers with higher salinity resistance and high capacity are directly used for separation and purification. The technological flow of the invention has the advantages of simple purification program step, uniqueness, low costs and convenient industrialization.