113 results about "Mutant cell" patented technology

Methods and vectors (both DNA and retroviral) are provided for the construction of a Library of mutated cells. The Library will preferably contain mutations in essentially all genes present in the genome of the cells. The nature of the Library and the vectors allow for methods of screening for mutations in specific genes, and for gathering nucleotide sequence data from each mutated gene to provide a database of tagged gene sequences. Such a database provides a means to access the individual mutant cell clones contained in the Library. The invention includes the described Library, methods of making the same, and vectors used to construct the Library. Methods are also provided for accessing individual parts of the Library either by sequence or by pooling and screening. The invention also provides for the generation of non-human transgenic animals which are mutant for specific genes as isolated and generated from the cells of the Library.

Methods and vectors (both DNA and retroviral) are provided for the construction of a Library of mutated cells. The Library will preferably contain mutations in essentially all genes present in the genome of the cells. The nature of the Library and the vectors allow for methods of screening for mutations in specific genes, and for gathering nucleotide sequence data from each mutated gene to provide a database of tagged gene sequences. Such a database provides a means to access the individual mutant cell clones contained in the Library. The invention includes the described Library, methods of making the same, and vectors used to construct the Library. Methods are also provided for accessing individual parts of the Library either by sequence or by pooling and screening. The invention also provides for the generation of non-human transgenic animals which are mutant for specific genes as isolated and generated from the cells of the Library.

The invention relates to methods for the stabilisation of cytokines and to cytokines generated by such methods. The methods of the invention involve mutating the amino acid sequence of a cytokine so as to remove solvent-exposed hydrophobic residues, and/or mutating the amino acid sequence of the cytokine so as to stabilise one or more secondary structure elements in the molecule. These steps have the effect of destabilsing intermediates that are formed during the folding process, relative to the stability of the cytokine in its naturally folded state, so increasing the yield of the cytokine as produced in vitro.

The present invention relates to a cell-based method for producing influenza virus vaccines by enriching the population of surface-bound α2,6-sialic acid receptors on a cell surface, such as on a Chinese Hamster Ovary (CHO) cell surface. The host cell therefore presents numerous binding sites to which an influenza virus can bind via its hemagglutinin spike protein and infect the host cell. In contrast to wild-type CHO cells, the surface of the mutated CHO cells of the present invention contains an enriched population of α2,6-sialic acid receptors which makes the inventive CHO cells highly susceptible to viral infection, and therefore safe, effective, and highly efficient cells for rapidly producing influenza vaccines.

The present invention relates e.g., to QM-7 or QT-6 cells comprising a heterologous mutant nicotinic α7 acetylcholine receptor and/or a nucleic acid encoding it, or a fragment or variant thereof. In a preferred embodiment, the mutant nicotinic α7 acetylcholine receptor subunit has a mutation in the M2 domain. QM-7 and QT-6 cells of the invention are useful for, e.g., assays such as high throughput assays that measure the influx of cations, such as Ca⁺⁺ ions, into a cell. Such assays can be used, e.g., to identify agents that modulate the expression and/or activity of a mutant cell-surface-expressed channel receptor (e.g., the nicotinic α7 receptor), and which thus modulate, e.g., among other functions, processes involved in the central nervous system, such as learning and memory.

Novel murine polynucleotides are disclosed that individually identify novel genes into which a retroviral gene trap vector has integrated. Additionally, novel mutated murine ES cells are described that stably incorporate retroviral gene trap constructs into the specifically identified genes. The novel genes and cells thus defined are useful in functional genomic analysis, and in the discovery and development of new therapeutic and diagnostics agents and methods.

Methods and vectors (both DNA and retroviral) are provided for the construction of a Library of mutated cells. The Library will preferably contain mutations in essentially all genes present in the genome of the cells. The nature of the Library and the vectors allow for methods of screening for mutations in specific genes, and for gathering nucleotide sequence data from each mutated gene to provide a database of tagged gene sequences. Such a database provides a means to access the individual mutant cell clones contained in the Library. The invention includes the described Library, methods of making the same, and vectors used to construct the Library. Methods are also provided for accessing individual parts of the Library either by sequence or by pooling and screening. The invention also provides for the generation of non-human transgenic animals which are mutant for specific genes as isolated and generated from the cells of the Library.

The present invention relates to methods of obtaining a mutant cell from a filamentous fungal parent cell, comprising: (a) obtaining mutant cells of the parent cell; (b) identifying the mutant cell which exhibits a more restricted colonial phenotype and/or a more extensive hyphal branching than the parent cell; and (c) identifying the mutant cell which has an improved property for production of a heterologous polypeptide than the parent cell, when the mutant and parent cells are cultured under the same conditions.

The invention provides a 2-aminopyridine derivative containing a 2-pyridone ring side chain or an enantiomer of the 2-aminopyridine derivative. The target compound is obtained mainly by taking 2-aminopyridine as a parent nucleus to be coupled with a 4-bromo-2-pyridone ring. Experiments prove that the 2-aminopyridine derivative has a significant proliferation inhibition effect on tumor cells Karpas 299 (NPM-ALK positive cell lines) related to the ALK tyrosine kinase activity, NCI-H2228 (EML4-ALK positive cell lines), SKN-BE2 (ALK gene amplification cell lines) and SH-SY5Y (ALK F1174 mutant cell lines) and can be prepared into corresponding anti-tumor-cell drugs. The 2-aminopyridine derivative has a general structural formula I (please see the formula in the description).

The invention provides a method of isolating desiccation resistant cells which retain good viability over long periods of time at room temperature for a variety of applications. In one embodiment, competent cells for DNA transformation are made from selected bacterial cells which are storage stable at room temperature.

Provided are compounds for inhibiting Snail-p53 binding and therapeutic agents for cancer including the compounds as an effective component. The Snail-p53 binding inhibitors induce expression of p53 in K-Ras mutant cell lines, thereby enabling effective treatment or prevention of K-Ras mutant cancer, such as, pancreatic cancer, lung cancer, cholangioma, and colon cancer, of which diagnosis or treatment is not easy.

The invention discloses the effect of a novel mutant human cytokine and fusion protein thereof on treatment of metabolic diseases. The human cytokine with mutant expression in mammalian cells is protein as shown in SEQ ID No. 3 in a sequence table. According to the invention, a wild human cytokine gene is subjected to mutation and N-terminal reconstruction, and the fusion protein contains a human cytokine mutant and the Fc segment (CH2-CH3) of human IgG4. The mammalian cell-expressed recombinant cytokine fusion protein has good activity in reducing blood sugar; in particular, the fusion protein has the advantages of high stability, long half life, etc.; and the fusion protein can well control fluctuation of blood sugar and stably maintain blood sugar at a normal level within 24 h.

The invention relates to the field of gene engineering and cytology, in particular to an AAV receptor knocking out method, an HEK293 cell strain with an AAV receptor being knocked out and an application. The knocked-out AAV receptor is KIAA0319L. The knocking out method comprises the following steps: establishing a sgRNA carrier of a targeted AAVR genome sequence; carrying out transfection for a 293 T-cell, and culturing to obtain each clonal cell; carrying out Suveyor gene mutation analysis test; selecting AAVR mutant monoclone; then selecting an AAVR mutant cell strain with high yield of AAV; identifying a mutation condition of other potential mutant site; and enlarging the cell strain without other mutant sites, and establishing a library for production. The final yield of the AAV produced by the HEK293 cell strain of the knocked-out AAV receptor is about 50 percent higher than the yield of the conventional HEK293.

The invention provides a preparation method of high-mutation urine stem cells in mitochondria mt.3243A > G mutation crowds. The method comprises the following steps: collecting urine cells of mitochondria mt.3243A > G mutant population; obtaining a sediment red tube by adopting a mode of centrifuging and adding a PBS to blow, beat and mix uniformly, obtaining P1-generation high-mutation urine stemcells are obtained, and obtaining the mitochondria mt.3243A > G mutation urine stem cells with the high mutation rate of 95% or above through digestion passage and mutation rate detection. The methodhas the advantages that the method can be obtained in a non-invasive mode, and the method can be obtained regardless of the gender, age and health condition of the patient. As a primary cell, the cell line maintains the basic properties of the original cell. The invention provides a cell application platform for researching the pathogenesis of mt.3243A > G mutant cells in the future and screeningnew drugs aiming at mt.3243A > G.

Novel mutated mammalian cells are provided that have been characterized by identifying the sequence of the genes that have been mutated. Preferably, novel mutated cells are murine ES cells that stably incorporate retroviral gene trap constructs in the specifically identified genes. The novel mutated cells and animals are useful in functional genomic analysis, and in the discovery and development of new therapeutic and diagnostics agents and methods.

The present invention relates to a conditionally inducible site-directed mutant cell, comprising a mutated allele of a gene; wherein said allele comprises a mutation that was introduced by using a suitable mutagenesis technique, a rescue allele of said mutated gene that can be conditionally inactivated, wherein said mutation in said mutated allele of said gene interferes with survival and/or causes, an adverse phenotype, such as temporal and/or local phenotypes, such as cell cycle-specific, cell-type specific, tissue-specific, protein-expression specific, tissue-development specific, organ-specific, organ-development-specific and/or embryonic lethal phenotypes. According to further aspects thereof, the present invention relates to a conditionally inducible site-directed mutant cell culture, tissue, organ, or non-human embryo, comprising a cell and a respective non-human organism, in particular a genetically deficient or Knock-outmammal, -rodent, -nematode, -fish, -plant or -insect. Finally the invention provides a method for inducible site-directed mutagenesis through conditional gene rescue, either in vitro or in vivo.

The invention provides a pyrrolo [2, 3-d] pyrimidine derivative targeting EGFR mutation as well as a preparation method and application thereof, and belongs to the field of chemical medicines. The derivative is a compound shown as a formula I, or a salt thereof, or a stereoisomer thereof. The compound disclosed by the invention is low in toxicity to normal cells, has an obvious inhibition effect on a lung cancer cell line, particularly has good selectivity on EGFR mutant cells HCC827, and has an obvious inhibition effect; meanwhile, the compound provided by the invention can effectively inhibit phosphorylation of EGFR. In addition, the compound provided by the invention has good inhibitory activity and selectivity on mutant EGFR. The compound disclosed by the invention can be used for treating lung cancer, particularly non-small cell lung cancer, has a relatively strong inhibition effect on EGFR mutant lung cancer, and is relatively low in toxicity; the compound can also be used for preparing tyrosine kinase inhibitors, especially EGFR phosphorylation inhibitors, and has good application prospects.

The invention relates to a method for screening biallelic mutant cell lines by Cas12a protein, is mainly used for screening biallelic mutant cell lines after CRISPR/Cas9 editing and solves the technical problem that biallelic mutant cell lines are difficult to screen out. A nucleic acid detection technology developed on the basis of Cas12a protein (including all bacterial sources) can screen biallelic mutant positive cells after CRISPR/Cas9 editing simply, efficiently and stably, and the method can partially replace a sequencing method and greatly reduces the screening cost.