68 results about "Transformed cell" patented technology

The present invention relates to a method, preferably a computer-implemented method, and to an apparatus for generating at least one intermediate cellular state of a cellular state evolution from an initial cellular state to a corresponding advanced cellular state. The present invention also relates to a computer program for generating at least one intermediate cellular state and to a non-transitory computer readable data medium storing the computer program. Moreover, the present invention relates to a method and to an apparatus for training a model for generating at least one intermediate cellular state of a cellular state evolution from an initial cellular state to a corresponding advanced cellular state. The present invention also relates to a computer program for training a model for generating at least one intermediate cellular state and to a non-transitory computer readable data medium storing the computer program. Furthermore, the present invention relates to a method for diagnosing a disease and forecasting patients' future clinical outcome making use of the method for generating at least one intermediate cellular state. Moreover, the present invention relates to an in-vitro method for transforming the identity of a cell.

The invention belongs to the technical field of cell biology, and particularly relates to a preparation method of tumor enhanced targeting and therapeutic function extracellular vesicles and a chip device. The invention firstly provides an integrated chip device which is provided with a cell screening module and a functional preparation module and comprises a plurality of functional layers, wherein one functional layer is provided with a cell evolution screening area with gradient for cell screening, and the other functional layer is provided with a cell electric remodeling area for cell functionalization transformation. The invention innovatively proposes that tumor cells with strong metastasis ability are screened based on culture gradient, the tumor cells with stronger invasion ability are distinguished by utilizing spatial segmentation, then the screened cell subpopulation is transferred to a cell electric remodeling area in combination with overturning operation of the chip, and the cell subpopulation is transferred to the cell electric remodeling area in a manner of focusing an electric field through nanopores. And meanwhile, therapeutic molecules are efficiently delivered to realize cell transformation, so that the transformed cells can enhance the output of extracellular vesicles with a therapeutic function.

The present invention describes mRNA usage improving and / or translation-enhancing nucleic acid sequences, nucleic acid constructs comprising such sequences, and host cells comprising such nucleic acid constructs. The invention further pertains to a method for expressing a protein of interest in a cell or organism using such nucleic acid sequences, as well as their uses for increasing integration of such nucleic acid construct into a genome, for enhancing mRNA usage and / or translation of a recombinantly expressed polypeptide, and for increasing the number of transformants upon transformation of a cell with such nucleic acid construct.

A nucleic acid construct is disclosed which is removable after transformation. Methods of using same are disclosed as well.

Chemical agents are added to standard production methods for recombinant adeno-associated viral vector to alter intracellular trafficking pathways in producer cells. This achieves, per producer cell, the synthesis of more genome containing vector particles and particles that are better transducing agents. Vector so produced also has: a greater propensity for export out of the producer cell, a more gradual transduction of infected cells, an enhanced ability for integration into the genome of cells that are coinfected by the adeno-associated virus, and an increased probability of transforming cells with multiple vectors.

Provided are methods of selecting a transformed cell, including methods comprising delivering an inhibitor of a protein to a plurality of cells, exposing the plurality of cells to a compound, wherein the protein (if present and active) converts the compound to a toxic compound, and selecting viable cells from the plurality of cells not killed by exposure to the compound.

PIGGYBAC® transposases engineered to increase stability in a cell. The engineered PIGGYBAC® transposases are useful for stably transforming cells, cell line development, genome modification, and improving titer of recombinant proteins, among other uses.

The application discloses a food-like lactobacillus double-plasmid multi-gene editing system based on CRISPR / Cas9 technology and application thereof. The food-like lactobacillus double-plasmid multi-gene editing system comprises (1) a plasmid pCRI01 comprising the following elements: a Cas9 protein coding gene; a temperature-sensitive replicon pSC101; an sgRNA expression module targeting the replicon of the plasmid pCRI02, which is controlled by an inducible promoter Pnis; an ampicillin resistance; and (2) a plasmid pCRI02 comprising the following elements: at least one sgRNA expression module driven by a P23 promoter; a lambda-Red recombinase system comprising Exo, Beta and Gam genes; an erythromycin resistance marker gene; and a kanamycin resistance gene. The multi-gene editing system provided by the application is a small-size pCRI01 and pCRI02 double-plasmid system, and the number of transformed cells after resistance screening is significantly improved; and the gene editing efficiency mediated by the pCRI plasmid is greatly improved with the assistance of the lambda-Red recombinase system.

A method of producing a transformed cell or plant body of maize of the present invention includes overexpressing, in a cell or plant body of maize, 1) a nucleic acid encoding an amino acid sequence of SEQ ID NO: 2, or a nucleic acid encoding a polypeptide including an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 2, the polypeptide having a function of promoting cell division of maize, or 2) a nucleic acid encoding an amino acid sequence of SEQ ID NO: 15, or a nucleic acid encoding a polypeptide including an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 15, the polypeptide having a function of promoting cell division of maize, the overexpressing being controlled by a promoter including a 35S promoter of Cauliflower mosaic virus.

Provided are a μ-conotoxin peptide CnIIIC mutant and the use thereof. Further provided are a nucleic acid construct of the μ-conotoxin peptide CnIIIC mutant, and an expression vector and transformed cell thereof, and the use thereof. The μ-conotoxin peptide CnIIIC mutant has an enhanced transdermal penetration ability, and can effectively penetrate the skin barrier to directly act on nerves and muscle tissues in the underlying layer of skin. By means of the unique physical and chemical properties, the mutant can realize a local pharmaceutical effect within a relatively short period of time, thereby greatly improving the bioavailability of a drug.

The invention belongs to the field of plant genetic engineering, and particularly relates to application of a WIND4 gene to improvement of genetic transformation efficiency of taxus chinensis. The WIND4 gene is constructed to a plant expression vector containing a Ruby report system, taxus chinensis cells are transformed through an agrobacterium-mediated method, on the premise that cell growth and development are not affected, rapid screening of positive cells can be achieved, division of transformed cells is promoted at the same time, and therefore transformation efficiency is improved. According to the invention, the synergistic interaction effect of WIND4 and Ruby is utilized, so that the defect of weak regeneration capacity of taxus chinensis cells is overcome. Compared with an empty vector control group, the WIND4 and Ruby co-expression can improve the transformation efficiency of taxus chinensis by about 1.4 times; compared with the traditional method, the conversion system disclosed by the invention is shorter in conversion period and simpler and more convenient to operate.

The invention discloses an application of circular RNA (Ribonucleic Acid) in tumor diagnosis and / or treatment. According to the present invention, high-throughput sequencing is adopted to detect the disordered expression circRNA in the metastasis cells, then a stable lung cancer cell model for overexpressing and knocking down the target circRNA is constructed, CCK-8 and transwell migration, invasion and other experiments are performed, and the cyclic structure of the circDCP2 and the up-regulation expression of the circDCP2 in the lung cancer cells are confirmed and verified, the result shows that knock-down of the circ DCP2 has a remarkable inhibition effect on the vicious transformation of BEAS-2B cells induced by the CBNPs. The circDCP2 can be used as a reliable biomarker for early lung cancer diagnosis and disease treatment.

The present disclosure provides a hotspot useful for transformation of animal cells by TI (Targeted Integration). The hotspot of the present disclosure was found near LOC103164262 in the CHO cell genome. Alternatively, the present disclosure relates to transformed cells into which an exogenous DNA has been introduced into said hotspot, and a method of producing a polypeptide encoded by the DNA by culturing the cells.

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.

A monoclonal antibody is provided which binds to a human CC chemokine receptor 1 (CCR1) and inhibits activation of the human CCR1, or an antibody fragment thereof. The monoclonal antibody binds to an extracellular region of a human CCR1 and inhibits activation of the human CCR1 by a human CC chemokine ligand 15 (CCL15). An antibody fragment thereof, a hybridoma producing the antibody, a nucleic acid having a nucleotide sequence encoding the antibody or the antibody fragment, a transformant cell containing a vector containing the nucleic acid, a method for producing the antibody or the antibody fragment using the hybridoma or the transformant cell; a therapeutic agent and a diagnostic agent containing the antibody or the antibody fragment, and a method for treating and diagnosing a CCR1-related disease using the antibody or the antibody fragment are also provided.

This invention provides transformed cells for the production of transformed replica pigs for xenotransplantation. [Solution] This invention relates to a transformed replica pig from which the heterologous antigens GGTA1, CMAH, iGb3s, β4GalNT2, and β2M genes have been removed from a PERV Envelope C-negative base, and a method for producing the same. The transformed replica pig according to the present invention can overcome hyperacute and antigen-antibody-mediated immune rejection reactions and T-cell-mediated immune rejection reactions without causing the transfer of porcine endogenous retroviruses that occur in xenologous organ transplantation, and can be usefully utilized as a donor animal for interspecies organ and cell transplantation.

The present invention relates to: an expression cassette comprising a scaffold / matrix attachment region (S / MAR); an expression vector for expressing a target protein, comprising the expression cassette; a transformant cell obtained by introducing the expression vector into a host cell; and a method for producing a target protein, comprising the step of culturing the transformant cell.

The present invention relates to a monoclonal antibody or an antibody fragment thereof that binds to an extracellular region of FCRL1, a hybridoma that produces the antibody, a nucleic acid comprising a nucleotide sequence that encodes the antibody or the antibody fragment thereof, a cell transformed with a vector comprising the nucleic acid, a method for producing the antibody or the antibody fragment thereof using the hybridoma or the transformed cell, an antibody-drug conjugate comprising the antibody or the antibody fragment thereof, a therapeutic agent and a diagnostic agent, each of which comprises the antibody or the antibody fragment thereof, and a therapeutic method and a diagnostic method for a disease associated with FCRL1 using the antibody, the antibody fragment thereof, or the antibody-drug conjugate comprising the antibody or the antibody fragment thereof.

Provided herein is a method for eliminating O-xylosylation modification of a recombinant protein, comprising constructing a mammalian cell having reduced O-xylosylation activity. The cell construction comprises: transfecting a cell with a CRISPR / Cas9 vector comprising an sgRNA sequence targeting a Xylt2 gene; and harvesting a transformed cell having reduced O-xylosylation activity via Xylt2 gene knockout.

The present invention provides a method for producing a target protein by culturing cells, the method being capable of stably maintaining a target plasmid without using an antibiotic. The present invention relates to a method for producing a target substance, the method including a step for culturing cells that are transformed with a target plasmid (A), wherein: each of the transformed cells includes the target plasmid (A) and a temperature-sensitive plasmid (B) and are obtained by a method that includes a step for culturing cells in each of which the function of an essential gene on the chromosome has been reduced at a temperature at which the temperature-sensitive plasmid (B) is eliminated; the target plasmid (A) includes the essential gene and a target substance gene and does not include an antibiotic-resistant gene; and the temperature-sensitive plasmid (B) includes the essential gene and the antibiotic-resistant gene.

The disclosure pertains to methods and compositions for the rapid and efficient transformation of plants. The disclosure further provides methods for producing a transgenic plant, comprising (a) transforming a cell of an explant with an expression construct comprising (i) a nucleotide sequence encoding a WUS / WOX homeobox polypeptide; (ii) a nucleotide sequence encoding a polypeptide comprising two AP2-DNA binding domains; or (iii) a combination of (i) and (ii); and (b) allowing expression of the polypeptide of (a) in each transformed cell to form a regenerable plant structure in the absence of exogenous cytokinin, wherein no callus is formed; and (c) germinating the regenerable plant structure to form the transgenic plant. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

As botulinum toxin has recently been attempted as a therapeutic agent for various indications, the demand for botulinum toxin in the medical and cosmetic fields has rapidly increased. However, there is no stable and highly reproducible cell-based assay method for measuring the potency of botulinum toxin. Since botulinum toxin is a very potent neurotoxin protein, the development of highly specific and sensitive cells is particularly required for accurate cell-based measurement of the potency of botulinum toxin. The present invention is directed to a neuronal cell line with altered gene expression. The transformed cell line according to the present invention has significantly increased sensitivity to botulinum toxin, and thus is expected to be very effectively used for cell-based determination of botulinum toxin activity or cell-based detection of botulinum toxin.

The present application provides a cell culture sheet. The cell culture sheet according to an embodiment of the present application is realized as including a fiber web having a three-dimensional network structure formed by accumulation of support fibers having an average diameter of 1.5 μm or less, a basis weight of 1 to 15 g / m 2 2; and a functional coating layer covering at least the support fibers exposed on one surface of the fiber web and having a function of promoting one or more of cell attachment, migration, proliferation, and differentiation. Thus, due to the high specific surface area and the surface morphology suitable for cells, cell attachment can be improved, the attached cells can be stably supported, cells can be cultured in a highly concentrated manner with high culture efficiency, and the cells can be cultured and recovered without forming a thin film or the like. In addition, compared to cells cultured by conventional culture, small-sized cells can be obtained, and thus, it is very advantageous to culture, proliferate, and renew fresh, healthy, and non-cell-transformed cells. Furthermore, excellent effects are obtained in differentiation and proliferation of stem cells differentiated into specific target cells.

A visual plasmid for replacing fluorescent proteins and visualization method for evaluating plant protoplasts transformation and transformation efficiency by replacing fluorescent proteins with visual plasmid. Genes expressed after the three kinds of plasmids are transformed into protoplasts metabolizes a substrate L-DOPA, causing protoplast cells to produce betacyanin or betaxanthin that are visible to the naked eye. The two pigments are visible to the naked eye under an optical microscope and may be used to intuitively determine whether a protoplast transformation experiment is successful. The plasmids can stably indicate successfully transformed cells. Their self-luminous intensities are superior to intensities of red fluorescent proteins tdtomato and EGFP, and can perfectly replace the role of the fluorescent proteins in evaluating protoplast transformation efficiency. This technology greatly simplifies the protoplast transformation experiment, gets rid of dependence on a fluorescent light source, and lowers the threshold for using the protoplast transformation technology.

Provided herein is a method of eliminating O-xyloylation modification of a recombinant protein, comprising constructing a mammalian cell having reduced O-xyloylation activity. The cell construction comprises the following steps: transfecting a cell by using a CRISPR / Cas9 vector containing an sgRNA sequence of a targeted Xylt2 gene; and harvesting the transformed cell with the O-xylosylation activity reduced and the Xylt2 gene knocked out.

The invention provides a method of treating a disease or disorder in a subject by inducing a TGF alpha immune response or by administering an anti-TGF-alpha antibody or a biologically active fragment thereof. The TGF-alpha immune response is induced using a TGF-alpha polypeptide or biologically active fragment, a vaccine, a genetic construct or a transformed cell, for example.

The present invention relates to a fusion protein in which a target protein and an Oct-1 protein-containing protein tag are linked, an expression structure comprising a nucleotide sequence encoding same, a recombinant vector including same, and a transformed cell including same.

The invention discloses a high-light-resistant algal strain high-speed screening method and system based on micro-fluidic single cell sorting, and relates to the field of bioengineering.The high-light-resistant algal strain high-speed screening method comprises the steps that haematococcus free cells in the logarithmic phase are taken and added into a double-fluorescence probe mixed solution, and incubation is conducted in a dark place to obtain a haematococcus solution; bBM sheath fluid and haematococcus fluid are introduced into a sample introduction area of the micro-fluidic chip, the BBM sheath fluid and the haematococcus fluid are focused by the sample introduction area and then enter a highlight stress area, highlight treatment is carried out to induce motile cells to be transformed into spores, and the transformed cells enter a detection area. Through in-situ high light stress and single cell continuous sorting of the micro-fluidic chip, the sorting efficiency is high, and the screening period is greatly shortened; three parameters of chlorophyll fluorescence, astaxanthin fluorescence and forward scattering light are adopted for cooperative detection, and a threshold value which is quantitatively set based on a control group is combined, so that the proportion of a false positive strain is reduced to be less than 5%, and the screening accuracy of a double-superior strain is greatly improved; the microfluidic chip is extremely low in single reagent consumption, greatly reduces the screening cost of single strains, and is easy for large-scale popularization and application.