91 results about "Transformed cell" patented technology

Methods and systems for performing a bioproduction process are described, the bioproduction process comprising the production of a transformed cell population. The methods include the steps of culturing a cell population at least partially embedded in a hydrogel matrix forming a three-dimensional structure, the hydrogel matrix also comprising a transforming agent.

The invention discloses an rhFN recombinant human chemical fibronectin process of a genetic engineering technology, and particularly relates to the technical field of genetic engineering, and the rhFN recombinant human chemical fibronectin process comprises target gene acquisition, gene cloning and vector construction, host cell selection and transformation, cell culture and screening, protein expression and optimization, and protein separation and purification. According to the rhFN recombinant human chemical fibronectin process of the genetic engineering technology provided by the invention, high-quality target genes are accurately obtained through technologies such as new-generation sequencing and gene synthesis, and the gene expression efficiency and stability are enhanced by virtue of optimized vector construction and regulatory elements; protein expression ability and specificity are improved in host cell selection and transformation, efficient culture and accurate screening are realized in cell culture and screening links, total yield is increased in a protein expression optimization link, purity and efficiency are improved in a protein separation and purification link, and product quality and safety are ensured through comprehensive quality control. The requirements in the fields of medical treatment and scientific research are expected to be better met.

The present invention relates to methods of synthesizing long-chain polyunsaturated fatty acids, especially eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, in recombinant cells such as yeast or plant cells. Also provided are recombinant cells or plants which produce long-chain polyunsaturated fatty acids. Furthermore, the present invention relates to a group of new enzymes which possess desaturase or elongase activity that can be used in methods of synthesizing long-chain polyunsaturated fatty acids. In particular, the present invention provides ω3 destaurases, Δ5 elongases and Δ6 desaturases with novel activities. Also provided are methods and DNA constructs for transiently and / or stably transforming cells, particularly plant cells, with multiple genes.

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.

To provide an expression construct for a protein for enhancing spines that can be introduced into neurons to increase a spine volume.SOLUTION: Provided are: [1] an expression construct for a protein for enhancing spines including a promoter for expression that functions in nerve cells, a domain of a Kalirin-7 protein required for at least spine enhancement, operably linked to the promoter, and a nucleotide sequence encoding a protein J1 that forms a heterodimer in the presence of a dimer-inducing compound; and [2] an expression construct for a protein for enhancing spines including a promoter for expression that functions in nerve cells, a domain of a PSD-95 protein required for the localization to at least a postsynaptic membrane, operably linked to the promoter, and a nucleotide sequence encoding a protein J2 that forms a heterodimer in the presence of a dimer-inducing compound.SELECTED DRAWING: None

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 herein is a promoter sequence consisting of a nucleic acid of between 12 and 300 nucleotides in length, wherein the nucleic acid comprises a polynucleotide having at least 90% identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and / or SEQ ID NO:6. Also provided are expression cassettes and vectors comprising the novel promoters described herein, wherein the promoter is operably linked to a preselected DNA segment encoding a protein or heterologous RNA transcript. Also provided are isolated transformed cells comprising the novel expression cassette or vector described herein. Methods are also provided for producing transformed cells and for treating a genetic disorder.

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 invention belongs to the field of biomedicine, and relates to an adeno-associated virus engineered with a targeting gene editing enzyme and a pharmaceutical composition thereof. Specifically, the present invention relates to a modified adeno-associated virus, a capsid protein of which is linked to a DNA cleavage enzyme. The present invention also relates to a transformed cell. The present invention can achieve in one step a knock-in of a gene fragment having a length up to 3.5-4K to a T cell and a lock-out of multiple genes from the T cell, and has good application prospects.

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.

Provided herein is a promoter sequence consisting of a nucleic acid of between 12 and 300 nucleotides in length, wherein the nucleic acid comprises a polynucleotide having at least 90% identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and / or SEQ ID NO:6. Also provided are expression cassettes and vectors comprising the novel promoters described herein, wherein the promoter is operably linked to a preselected DNA segment encoding a protein or heterologous RNA transcript. Also provided are isolated transformed cells comprising the novel expression cassette or vector described herein. Methods are also provided for producing transformed cells and for treating a genetic disorder.

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.