56 results about "Wild type cell" patented technology

A recombinant microbial cell is capable of producing at least one compound having structural Formula II from a carbon source:In Formula II, n=2-3; R1=H or COCH3 or CH2CH2COX with X=OH or O—; R2=CH3 or CH2CH2COX with X=OH or O— where the cell comprises a genetic modification to increase activity relative to its wild-type cell of E4 where E4 is a desferrioxamine or bisucaberin synthetase (EC 6.3.-.-) (E4i) capable of converting N5-aminopentyl-N-(hydroxy)-succinamic acid to desferrioxamine B or H or at least one other linear desferrioxamine or bisucaberin according to Formula II.

The invention discloses a tetra-heterocyclic compound as well as a preparation method and application thereof, and particularly discloses a compound as shown in a formula I, and a stereoisomer or pharmaceutically acceptable salt thereof. The tetra-heterocyclic compound provided by the invention has good proliferation inhibition activity on at least one of NCI-H358 cells, Capan-1 cells and AsPC-1 cells containing KRAS mutation, has relatively weak proliferation inhibition activity on KRAS wild type PC-9 cells, and is expected to treat and / or prevent KRAS mediated diseases.

The invention discloses a biaromatic ring amide compound and application thereof. The invention provides a compound as shown in a formula I or pharmaceutically acceptable salt thereof. The small molecule compound provided by the invention has a good inhibition effect on Pol theta enzyme and / or PARP1 enzyme, further has a relatively strong proliferation inhibition effect on DLD1BRCA2- / -cells, has an extremely weak proliferation inhibition effect on DLD1 wild-type cells and shows very good selectivity, and further, the compound provided by the invention can be used for overcoming drug resistance, reducing the dosage and reducing the toxic and side effects, and has a good application prospect. The purposes of expanding indications and improving the treatment effect are achieved. # imgabs0 #

The present invention relates to mutant enzymes with enhanced properties and methods of using such enzymes to oxidize organic compound substrates. More specifically, provided herein are mutations in cytochrome P450 enzymes that enhance their peroxygenase activity and / or alter product selectivity as compared to wild-type cytochrome P450 enzymes. Also provided are uses of such mutant cytochrome P450 enzymes, e.g., for biocatalytic oxidation of carbon-hydrogen bonds by a peroxygenase pathway, or for oxidation of organic compound substrates.

To provide, in one embodiment, a method of using human-induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery.SOLUTION: In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.SELECTED DRAWING: None

The invention discloses a chimeric antigen receptor targeting EGFRvIII and application of the chimeric antigen receptor. The chimeric antigen receptor sequentially comprises an EGFRvIII targeting nucleic acid aptamer, a transmembrane domain and an intracellular signal domain from an N end to a C end, the EGFRvIII targeted nucleic acid aptamer is an EGFRvIII specific DNA aptamer obtained on the basis of SELEX (systematic evolution of ligands by exponential enrichment) screening, can only be combined with an extracellular domain of the EGFRvIII, and is not combined with an EGFR (epidermal growth factor receptor) wild type; the intracellular signal domain comprises a costimulatory signal domain and an activation signal domain, the C end of the costimulatory signal domain is connected with the N end of the activation signal domain, and the EGFRvIII specific nucleic acid aptamer screened based on SELEX is adopted as a targeting domain, can only be specifically combined with the extracellular domain of EGFRvIII positive tumor cells, and is not subjected to cross combination with EGFR wild cells and normal cells, so that the EGFRvIII specific nucleic acid aptamer can be used for detecting EGFRvIII positive tumor cells, and the EGFRvIII specific nucleic acid aptamer can be used for detecting EGFRvIII positive tumor cells. The problem of off-target killing caused by the fact that a common antibody scFv fragment of a traditional chimeric antigen receptor (CAR) is easily combined with a wild type EGFR is fundamentally avoided.

T lymphocytes play a key role in the immune response and their functions are intimately linked to metabolic programs. During immune responses, T cells undergo a metabolic reprogramming notably characterized by an increased aerobic glycolysis. Using a quantitative phosphoproteomic approach, the inventors have identified a new transcription factor called Foxk1 as being highly phosphorylated in T cells upon T Cell Receptor (TCR) engagement. The results also indicate that Foxk1 phosphorylation and nuclear translocation is dependent of the AKT-mTOR kinase activities. Using T-cell specific Foxk1 deficient mice (Foxk1- / -), we demonstrated that Foxk1 is required for full T cell activation. Foxk1-deficient T cells exhibited reduced proliferation and cytokine secretion following TCR stimulation. Furthermore, T cells from Foxk1- / - mice were less prone to acquire an effector like phenotype than wild-type cells when challenged in vivo. Conversely, Foxk1 overexpression in T cells enhanced their effector functions in a TCR-dependent manner. In CD8+ T cells, this effect also results into enhanced cancer cell killing capacity in vitro, and improved tumor rejection in vivo. Altogether, these results indicated that Foxk1 is a major regulator of T cell metabolism and thus, of T cell effector functions. Thus, the present invention relates to host immune cells engineered to overexpress a Foxk1 polypeptide and their use of the treatment of cancer.

The present invention discloses, in one embodiment, a method of using human induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery⋅. In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.

This invention relates to the biotechnological production of ω-functionalized carboxylic acids and their esters. Microbial cells are provided for the production of at least one ω-functionalized carboxylic acid ester from at least one alkane, wherein the cells are genetically modified relative to wild-type cells to increase the expression of: (i) an enzyme E1 capable of converting the alkane to the corresponding 1-alkanol; (ii) an enzyme E2 capable of converting the 1-alkanol of (i) to the corresponding 1-alkanal; (iii) an enzyme E3 capable of converting the 1-alkanal of (ii) to the corresponding alkanoic acid; (iv) an enzyme E4 capable of converting the alkanoic acid of (iii) to the corresponding alkanoic acid ester; and (iv) an enzyme E5 capable of converting the alkanoic acid ester of (iv) to the corresponding ω-hydroxy-alkanoic acid ester, and wherein the cells do not contain an increase relative to wild-type cells of the expression of the following enzymes selected from the group consisting of: 20 -E 24 Genetic modification of expression of at least one of them: -E 20 Acyl-ACP thioesterase, -E 21 Acyl-CoA thioesterase, -E 22 Acyl-CoA:ACP transacylase, -E 23 Polyketide synthase, and -E 24 Hexanoic acid synthase.

This disclosure relates to formulations comprising one or more cells, wherein the cells of the formulation are modified to conditionally express: (i) increased levels of one or more immune checkpoint proteins compared to corresponding wild-type cells; (ii) decreased levels of one or more HLA-I proteins compared to corresponding wild-type cells; or a combination of (i) and (ii). This disclosure further relates to methods and constructs for producing said cell formulations, and methods for administering said cell formulations to subjects in need.

The invention belongs to the field of gene engineering, and particularly relates to construction of an RNF31 gene knockout cell line and application of the RNF31 gene knockout cell line as a foot-and-mouth disease virus vaccine production cell line. The method comprises the following steps: firstly, designing two sgRNA sequences at an RNF31 exon by utilizing a CRISPR / Cas9 technology, and connecting with a pX459-puro vector to construct a recombinant plasmid; the method comprises the following steps: transfecting a PK-15 cell with pX459-RNF31-sgRNA, and screening under the action of puromycin, so as to obtain the RNF31 gene knockout cell line. Compared with a wild type cell, the RNF31 gene knockout cell line remarkably promotes FMDV replication and can be used as a production cell line of a foot and mouth disease virus vaccine; and data support is provided for further researching a mechanism of inhibiting FMDV replication by RNF31.

This invention discloses the application of T cells overexpressing PRDM16 in the preparation of anti-tumor or infectious disease drugs. Through experiments, this invention verifies that overexpression of PRDM16 enhances the in vivo and in vitro activity of T cells. It was found that in vitro overexpression of PRDM16 significantly enhances the killing effect of T cells on target cells. In vivo reinfusion of PRDM16 has stronger anti-tumor and anti-infection functions compared to wild-type T cells. Therefore, it can be used in malignant tumors and infectious diseases to enhance the efficacy of T cell adoptive therapy and solve the defect of T cell adoptive therapy where the killing power of target cells is suppressed due to environmental influences.

The invention discloses an application of a poultry IRF2 gene, and the poultry IRF2 gene is used for inhibiting the replication of Mycoplasma gallisepticum (MG) in host cells and the expression of virulence protein, so that the poultry chronic respiratory disease (CRD) caused by MG infection can be resisted. According to the application disclosed by the invention, the IRF2 gene in the chicken macrophage HD11 is interfered by utilizing a gene silencing technology, after MG challenge, the replication level and virulence protein expression of intracellular MG are obviously lower than those of wild type cells, inflammatory response and cell apoptosis of HD11 are obviously inhibited, and cell proliferation is promoted. The overexpressed IRF2 gene in the cell, the replication of the MG in the cell and the expression of the virulence protein are obviously enhanced, which indicates that the IRF2 gene is a susceptible gene of the MG, and the cell has an MG resistance phenotype after being knocked down, so that the replication of the MG and the expression of the virulence protein can be effectively inhibited. Therefore, the IRF2 gene can be used as a new target for development of vaccine drugs for inhibiting MG replication and research on disease resistance breeding.

The invention belongs to the technical field of medicines, and particularly relates to application of a ciclopirox compound in a medicine for treating cancers induced by KRAS G12D mutation, and the cancers comprise pancreatic ductal adenocarcinoma and colorectal cancer. The compound can obviously inhibit the activity of KRAS mutant pancreatic cancer cells, has weak inhibition on KRAS wild type cells, and shows potential targeted therapy advantages. Animal experiment results show that the ciclopirox compound provided by the invention can effectively inhibit tumor growth and down-regulate phosphorylated STK31 protein expression. The composition is clear in component and simple and convenient to prepare, and provides a new strategy for overcoming the drug resistance of pancreatic cancer.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a TP63 gene in improving the expression quantity of exogenous recombinant protein, and the nucleotide sequence of the TP63 gene is shown as SEQ ID NO.1. According to the invention, the TP63 overexpression vector is creatively applied to construction of a recombinant protein expression system, the TP63 overexpression vector is transfected into CHO cells to construct a CHO cell line for overexpressing TP63, and the CHO cell line can significantly improve the expression level of TP63 protein. Meanwhile, the CHO cell line is used as a host cell for recombinant protein expression, compared with a wild type CHO cell line, the expression quantity of recombinant protein can be increased, and the problem that the expression level of antibody expression quantity in an existing CHO cell expression system is low is effectively solved.

The invention belongs to the technical field of biology, and particularly relates to a VP2 protein mutated O-type foot-and-mouth disease recombinant virus as well as preparation and application thereof. The 88th lysine residue of O-type FMDV-VP2 protein is mutated into arginine residue by using a reverse genetic manipulation technology to successfully rescue the O-type foot-and-mouth disease recombinant virus strain rO-VP2K88R, and compared with an O-type foot-and-mouth disease wild type virus strain rO-WT rescued by using the reverse genetic manipulation technology, the recombinant virus strain provided by the invention has the advantages that the recombinant virus strain has the advantages that the yield is high; the growth replication capacity in wild type PK-15 cells is remarkably reduced, and the pathogenicity to suckling rats is also remarkably reduced. Therefore, the obtained O-type foot-and-mouth disease recombinant virus strain rO-VP2K88R possibly has certain immunogenicity and is expected to become a vaccine candidate strain, and a basic research material is provided for researching the foot-and-mouth disease vaccine candidate strain.

The invention relates to a bioengineering technology, and aims to provide an IPEC-J2 cell strain for stably expressing a PED virus nucleocapsid protein truncated fragment and application of the IPEC-J2 cell strain. The cell strain is preserved in the China Center for Type Culture Collection, the preservation name is a porcine small intestine epithelial cell line R58-WT-IPEC, the Latin name is Sus scrofa, and the preservation number is CCTCC NO: C202425. The invention discloses a key effect of the PEDV N protein in a virus replication stage, and provides application of the PEDV N protein as a target spot in regulation and control of PEDV infection. Compared with a wild-type IPEC-J2 cell line, the cell line provided by the invention has a remarkable promotion effect, can solve the problem of poor proliferation of PEDV (porcine epidemic diarrhea virus) in the IPEC-J2 cell line, and further improves the application possibility of the IPEC-J2 cell line in virus separation and propagation.

Targeting the ability of T cells to enhance aerobic glycolysis upon TCR stimulation or glucose supplementation represents an attractive strategy for the treatment of autoimmune diseases. Using a quantitative phosphoproteomic approach, the inventors have identified a new transcription factor called Foxk1 as being highly phosphorylated in T cells upon T Cell Receptor (TCR) engagement. The results also indicate that Foxk1 phosphorylation and nuclear translocation is dependent of the AKT-mTOR kinase activities. Using T-cell specific Foxk1 deficient mice (Foxk1- / -), the inventors demonstrated that Foxk1 is required for full T cell activation. Foxk1-deficient T cells exhibited reduced proliferation and cytokine secretion following TCR stimulation. Furthermore, T cells from Foxk1- / - mice were less prone to acquire an effector like phenotype than wild-type cells when challenged in vivo. Altogether, these results indicated that targeting can be envisioned to reduce auto-immune reactions. Thus, the present invention relates to the use of Foxk1 inhibitors for the treatment of inflammatory autoimmune diseases.

Provided are a mesenchymal stem cell (MSC) with an enhanced targeted tumor infiltration ability, and the use thereof. The MSC with an enhanced targeted tumor infiltration ability comprises a polynucleotide encoding an NKG2DL-targeting chimeric antigen receptor. The provided MSC with an enhanced targeted tumor infiltration ability can not only normally overexpress the NKG2DL-targeting chimeric antigen receptor, but also has a better solid tumor targeting property, thus being able to effectively break through solid tumor microenvironments and enter the interior of tumors. Compared to wild-type MSC cells, the MSC has a better retention ability at the tumor site, which enhances the survival ability thereof at the tumor site, thereby enabling the MSC to exert a more long-acting and sustained tumor inhibitory effect at the tumor site and further promoting the use of the MSC in the field of solid tumor treatment.

The invention provides an anti-leukemia small molecule compound TIP-20 targeting FLT3 and application of the anti-leukemia small molecule compound TIP-20, and belongs to the technical field of biological medicine. The invention develops a novel FLT3 kinase inhibitor TIP-20, which can enhance the antigen presentation effect of FLT3-ITD leukemia cells, can be combined with FLT3 protein, stabilizes the structure of FLT3, and further inhibits FLT3 phosphorylation and downstream pathway activation. The TIP-20 has an obvious inhibiting effect on the growth of leukemia cell strains, and can inhibit the growth of leukemia cells with FLT3 mutation by inhibiting phosphorylation of FLT3, promoting cell apoptosis and cycle arrest. Moreover, FLT3 wild type cells have good tolerance to TIP-20, have a wide therapeutic window with FLT3-ITD mutated acute myelogenous leukemia cells, and are expected to bring a better choice for treatment of FLT3 mutated AML patients.

The invention provides application of overexpressed SLC29A1 in enhancing internal and external activities of T cells, the killing effect of the T cells on target cells is remarkably enhanced through in-vitro overexpressed SLC29A1, in-vivo feedback has stronger anti-tumor and anti-infection functions compared with wild type T cells, and the T cells can be used for preparing drugs for resisting malignant tumors and infectious diseases and have good application prospects. And the curative effect of T cell adoptive therapy is enhanced.

The invention belongs to the technical field of cell engineering, and particularly relates to application of a TRAF3 gene and a TRAF3 gene knockout pig kidney cell line in preparation of a medicine for preventing and / or treating foot-and-mouth disease, and the Gene ID of the TRAF3 gene is 100156979. According to the invention, two sgRNAs are designed for a pig TRAF3 gene, the sgRNAs are constructed to a gene editing vector, and PK-15 cells are transfected, so that a TRAF3 gene function-deleted cell line is obtained. Compared with a wild type cell, the knockout cell line can significantly promote the replication of FMDV. It is shown that the TRAF3 gene plays an important regulation role in the host anti-FMDV replication process, and a new drug target is provided for preventing or treating FMDV. The TRAF3 gene knockout cell line disclosed by the invention has great scientific significance on wide application of a gene editing technology, prevention of virus transmission and guidance of a reverse genetic system to optimize vaccine design.

The invention discloses an application of a ZFYVE27 gene or an encoded protein thereof in regulating and controlling ASFV resistance of a host, and belongs to the fields of molecular biology and prevention and treatment of animal infectious diseases. A ZFYVE27 gene knockout or silence cell model is constructed by using a CRISPR / Cas9 gene editing technology and an RNAi technology, the gene knockout cells show significant resistance to ASFV infection, the proliferation speed of ASFV on the gene knockout cells is significantly lower than that of wild cells, and it is indicated that the ZFYVE27 gene plays an important role in regulating and controlling host virus resistance. The antiviral regulation function of the ZFYVE27 gene is identified from the cellular level, it is found for the first time that the gene can be used as a potential target for gene editing cells and animal design to be applied to disease-resistant breeding and drug research and development, and the gene has important significance for reducing drug abuse and avoiding epidemic situations.

Provided herein are genetically modified T cells that exhibit increased proliferation compared to wild-type T cells when stimulated, methods of generating such T cells, and methods of using the T cells for the treatment of a disease such as cancer.

The invention provides mesenchymal stem cells capable of enhancing tumor targeted infiltration capacity and application of the mesenchymal stem cells. The mesenchymal stem cell capable of enhancing the tumor targeted infiltration capacity comprises polynucleotide of a chimeric antigen receptor for coding targeted NKG2DL. The mesenchymal stem cell capable of enhancing the tumor targeting infiltration capacity not only can normally overexpress the NKG2DL targeting chimeric antigen receptor, but also has better solid tumor targeting property, and can effectively break through the solid tumor microenvironment and enter the tumor. Compared with wild type MSC cells, the MSC cells have better retention ability in tumor sites, and the survival ability of the MSC cells in the tumor sites is enhanced, so that the mesenchymal stem cells can play a more long-acting continuous tumor inhibition role in the tumor sites, and the application of the MSC cells in the field of solid tumor treatment is further promoted.

The invention discloses an engineered cell with low immunogenicity as well as a preparation method and application of the engineered cell. The expression level of the B2M gene, the CIITA gene, the CD54 gene and the CD58 gene of the engineered cell is lower than that of a wild type cell or is not expressed, so that the sensitivity of the engineered cell to T cells and NK cells is reduced, and the application of the engineered cell in cell therapy is promoted.

The invention discloses an application of T cells overexpressing PRDM16 in preparation of drugs for resisting tumors or treating infectious diseases, experiments prove that overexpressing PRDM16 can enhance in-vivo and in-vitro activity of the T cells, and discovers that in-vitro overexpressing PRDM16 enables the killing effect of the T cells on target cells to be remarkably enhanced; compared with wild type T cells, in-vivo feedback has stronger anti-tumor and anti-infection functions, so that the T cells can be applied to malignant tumors and infectious diseases, the curative effect of T cell adoptive therapy is enhanced, and the defect that killing power is inhibited due to the fact that target cells are affected by the environment by the T cell adoptive therapy is overcome.

Disclosed herein are single chain dimeric cytokines having improved physiological activity over wild-type cytokine monomers. The single chain dimeric cytokines of the disclosure may be useful in the treatment of various diseases or disorders relating to inflammatory conditions or proliferative conditions. Also disclosed are delivery vectors or agents for delivering the single chain dimeric cytokines to a subject. Such delivery vectors or agents include cells, bacteria and bacteriophages.

The present invention belongs to the technical field of anti-tumor drug research, and specifically discloses a method for preparing a therapeutic effector CD8 + The application of drugs in T cell differentiation related diseases. Experiments have shown that in mouse tumor models, adoptive specific modification of ADAM17 CD8 + T cells compared with wild-type CD8 + T cells can significantly inhibit tumor growth, and the exhaustion of CD8 + The proportion of T cells decreased. + Modification of ADAM17 in T cells can effectively increase CD8 + T cells can clear bacteria and enhance the ability to resist infection.

The present invention discloses the application of the MDH2 gene and its inhibitors in resisting ZEA toxin, which relates to the field of biotechnology. In the present invention, a cell line lacking the MDH2 gene was obtained through CRISPR gene editing. Compared with the cells expressing the wild-type MDH2 gene, under the influence of ZEA, the cell viability was extremely significantly improved, and the apoptosis rate of the cells was extremely significantly reduced. In addition, inhibiting the MDH2 activity of wild-type IPEC-J2 cells with the LW6 inhibitor can also improve the tolerance of the cells to ZEA. Thus, knocking out, mutating or inhibiting the activity of the MDH2 gene can improve the ability of humans, animals and plants to resist ZEA. The present invention provides a potential solution to solve the adverse reactions caused by ZEA.