79 results about "Knockout animal" patented technology

The invention discloses application of a JMJD6-targeted reagent in preparation of a medicine for promoting myocardial cell proliferation. The medicine can promote myocardial cell proliferation after myocardial infarction and reduce the myocardial fibrosis scar area caused by myocardial infarction. AAV9 myocardial specific overexpression virus and JMJD6 myocardial specific knockout mice are utilized, and the positive effect of JMJD6 in promotion of P1 cardiac apex resection of newborn mice and regeneration and repair of injured hearts after myocardial infarction of adult mice is disclosed for the first time; the specific mechanism is that JMJD6 depends on the activity of histone demethylase, enrichment of active modification H4R3me2a and H3R2me2s in a PDK4 promoter region is removed, transcriptional expression of the H4R3me2a and the H3R2me2s is inhibited, an impaired heart energy substrate utilization mode is stimulated to be increased and converted from fatty acid oxidation energy supply to glycolysis oxidation energy supply, and then adult myocardial cell proliferation is effectively promoted. The regeneration and repair capability of the heart after myocardial infarction is greatly improved, and a new effective target spot is provided for clinical treatment of myocardial injury.

The invention belongs to the technical field of disease model construction, and particularly relates to a construction method of a chronic myelogenous leukemia mouse model with conditional human EZH2 overexpression and RUNX1 knockout. According to the invention, a chronic myelogenous leukemia mouse transgenic mouse model with conditional human EZH2 overexpression and RUNX1 knockout is successfully constructed, the model is induced to be converted from a chronic stage to a sudden change stage, and particularly, the model is a transgenic mouse model which is positive in Lyz2-CreERT2 / EZH2 / RUNX1 and carries BCR-ABL and SCL-tTA. It is proved that a human EZH2 conditional overexpression and RUNX1 knockout chronic myelogenous leukemia mouse transgenic mouse model has feasibility and importance for research on conversion from CML CP to BC samples, and a molecular mechanism for conversion from chronic myelogenous leukemia to a sudden change stage is revealed for research. And a new animal model and a new research idea are provided for understanding of disease progression and development of a new treatment strategy.

The present disclosure provides an immunodeficient or immune-impaired chimeric non-human animal with a deletion or impairment of adeno-associated virus receptor (AAVR), comprising human hepatocytes, methods for preparing the chimeric non-human animal comprising human hepatocytes and methods of utilizing the chimeric non-human animal comprising human hepatocytes to evaluate transduction efficiency of adeno-associated viruses (AAV), and determine mechanism of inhibition / modification of AAV transduction in human hepatocytes.

Provided in the present invention is an sgRNA targeting the mouse FLT3 gene. Further provided in the present invention is a method for constructing a mouse having a humanized immune system, the method comprising: injecting an AAV vector comprising a cytokine gene into a severely immunodeficient FLT3-gene-knockout mouse; optionally irradiating the FLT3-gene-knockout mouse; and injecting human hematopoietic stem cells into the FLT3-gene-knockout mouse.

The use of FcγRIIB as a mammalian mammary IgG transport receptor. By constructing FcγRIIB-knockout mice and FcγRIIB-knockout pigs, it is found that the ratio of serum and milk IgG concentrations of the FcγRIIB-knockout mice is significantly increased, that is, the relative concentration of milk IgG is reduced; and the FcγRIIB-knockout pigs have a significantly increased serum IgG content, but IgG is nearly undetectable in colostrum. Therefore, it has been proved for the first time that FcγRIIB is a receptor that mediates the trans-mammary transport of serum IgG to milk, and answers a long-term unsolved problem in the field of maternal passive immunity research. Furthermore, a new strategy is provided for increasing the IgG content of colostrum, thereby improving the early immunity and survival rate of neonatal domestic animals.

The application discloses an IGFBP-2 derived polypeptide and application thereof in preparation of an antidepressant, and relates to the technical field of biological medicines. The amino acid sequence of the polypeptide is Pro-Lys-Lys-Leu-Arg-Pro, the N terminal of which is acetylated and the C terminal of which is amidated, and the amino acid sequence is shown as SEQ ID NO:1. The polypeptide has a small molecular weight, can easily penetrate the blood-brain barrier, and has good stability after terminal modification. Animal experiments prove that the polypeptide can significantly improve the depressive behavior of Shank3 gene knockout mice, including improving the spatial cognitive ability, enhancing the social interaction and curiosity, and relieving the anxiety behavior. Cell experiments prove that the polypeptide can significantly improve the viability of hippocampal neuron cells from Shank3 gene knockout mice, and up-regulate the expression level of synaptic plasticity related proteins GluA1 and SYN1. The polypeptide can be prepared by a chemical synthesis method, has low cost, and can be used for preparing a medicine for preventing or treating depression (especially depression related to Shank3 gene mutation).

The application discloses application of DNA methyltransferase inhibitors in preparation of drugs for treating polycystic kidney disease. Preferably, the DNA methyltransferase inhibitor is decitabine or azacitidine, and the polycystic kidney disease is polycystic kidney disease caused or aggravated by mTOR activation. The application uses Tsc2 knockout mouse embryo fibroblasts MEF and Tsc2 kidney-specific knockout mice, finds that after treatment of the DNA methyltransferase inhibitor decitabine, cell proliferation is inhibited, polycystic kidney disease of the mice is relieved, and kidney function is partially recovered. The application provides a new and effective technical means for treatment of polycystic kidney disease.

The application discloses a novel metabolic marker for preparing a medicine for treating liver cancer and application thereof. By constructing an ALDH6A1 overexpression liver cancer cell line, it is found through detection that the level of methylcitrate in the cell is inversely proportional to the proliferation and migration rate of liver cancer cells. In Aldh6a1 knockout mice, an AKT / NRAS liver cancer model is constructed by high-pressure tail vein injection, and it is found that the content of methylcitrate in the serum is inversely proportional to the levels of ALT and AST in the serum of the mice and inversely proportional to the liver cancer load of the mice. The metabolite methylcitrate can not only effectively inhibit the proliferation of liver cancer cells alone, but also can enhance the inhibitory effect of sorafenib on the proliferation of liver cancer cells. The metabolite can effectively inhibit the formation of tumors, can be used as a metabolic marker for detecting liver cancer, and can more accurately and efficiently judge the severity of liver cancer; and can be used as a novel metabolite for treating liver cancer, and can improve the treatment effect of liver cancer.

The invention belongs to the technical field of medical biological research, and particularly relates to a construction method of a spontaneous emphysema mouse model. The invention relates to a method for constructing a spontaneous emphysema mouse model, which comprises the following steps: when a Muc1 gene whole body knockout mouse naturally senility for 3-18 months to form spontaneous emphysema and / or a Muc1 type 2 alveolar epithelial cell knockout mouse is 6-8 weeks old, injecting tamoxifen into the intraperitoneal cavity of the mouse to induce knockout. According to the invention, the Muc1 gene knockout mouse is used for constructing a spontaneous emphysema mouse model product for the first time; the related products comprise construction schemes, application transformation and the like of a spontaneous emphysema model generated by Muc1 whole body knockout mouse (Muc1- / -) along with age increase and a spontaneous emphysema model generated by Muc1 type 2 alveolar epithelial cell knockout (Muc1AT2- / -) mouse induced by intraperitoneal injection tamoxifen along with age increase. The invention proves that the spontaneous emphysema model is successfully constructed.

The invention discloses a method for constructing a neurodevelopmental disorder animal model based on central nervous system myelin sheath function change and application, and belongs to the technical field of biological engineering. An Msl2 gene conditional knockout mouse model is constructed by adopting a gene engineering technology, the space-time specific knockout of a second exon of the Msl2 gene in a specific cell type is realized through a Cre-LoxP recombinase system, and the exon encodes a key enzyme activity region for catalyzing ubiquitination. Model construction is based on central nervous system oligodendrocyte / myelin sheath dysfunction, the behavior phenotype of the model is similar to the behavior of a typical neurodevelopment disorder animal, a brand new perspective is provided for exploring an etiology mechanism, model mice can be prepared on a large scale by performing directional mating on the gene modified mice, and the development of the model is promoted. And consistency of different experiment batches and reliable reproduction of experiment data are ensured.

The invention discloses a construction method of an endothelial cell knockout Cdc42 gene aggravated bleomycin induced pulmonary fibrosis mouse model, which comprises the following steps of: mating a Cdc42 gene conditional knockout mouse with a mouse (Tie2-CreER) of which the endothelial cell specifically expresses tamoxifen induced Cre recombinase to obtain a double-transgenic mouse (Cdc42fl / fl-Tie2-CreER); the Cdc42 gene is specifically knocked out in vascular endothelial cells under the induction of tamoxifen, and then pulmonary fibrosis is induced by subcutaneous injection of bleomycin. The model shows aggravated pulmonary fibrosis phenotypes, including collagen deposition increase, alveolar structure damage and fibrosis-related protein expression up-regulation, and is high in construction success rate and good in repeatability. The invention also provides an application of the model in research of systemic sclerosis related interstitial lung disease pathogenesis and screening of anti-pulmonary fibrosis drugs, and an application of the Cdc42 gene as a drug target, and provides an accurate and reliable tool for pulmonary fibrosis research.

The application discloses a vitamin D binding protein gene conditional knockout mouse model and application thereof. It is found that the mouse with the vitamin D binding protein specifically knocked out in microglial cells can resist chronic stress-induced depressive behavior; it is indicated that the vitamin D binding protein in the microglial cells is a key factor for the occurrence and development of depressive behavior; by knocking out the vitamin D binding protein in the microglial cells, a significant effect of resisting depressive behavior can be achieved; the vitamin D binding protein in the microglial cells can be used as a drug target for screening drugs for treating depression, and the vitamin D binding protein inhibitor in the microglial cells can be used for preparing the drugs for treating depression. The application opens a new direction for in-depth research on depression and provides more choices for clinical treatment of depression, and has very important theoretical and practical significance.

The invention discloses a construction method and application of a DNAJB6a gene specific knockout mouse model, and belongs to the technical field of gene engineering. Comprising the following steps: S1, designing gRNA sequences aiming at ninth to tenth exon regions of the mouse DNAJB6 gene; s2, in vitro transcription is carried out to prepare Cas9mRNA and gRNA; s3, carrying out microinjection on Cas9mRNA and gRNA (guide Ribonucleic Acid) into a fertilized egg of the mouse; s4, transplanting the fertilized ovum after injection into the body of a pseudo-pregnant female mouse to obtain an F0-generation mouse; s5, screening positive F0-generation mice through genotype identification, and mating the positive F0-generation mice with the wild type mice to obtain F1-generation heterozygote mice; s6, the F1-generation heterozygote mice are matched to obtain F2-generation homozygote mice, and the DNAJB6a gene specific knockout mouse model is established. The construction method for constructing the DNAJB6a gene knockout mouse model, provided by the invention, is simple and convenient to operate, short in construction process time and stable in genotype, and a DNAJB6a gene knockout mouse can be effectively obtained.

The application belongs to the technical field of biological medicine, and particularly relates to application of SFRP1 in treatment of aortic aging drugs. SFRP1 can induce generation of vascular organoids, significantly antagonize vascular smooth muscle cell senescence induced by angiotensin II and bleomycin, enhance vascular smooth muscle cell activity, improve cell oxidative stress ability, reduce cell beta-galactosidase activity, reduce intracellular calcium deposition level, reduce phenotype conversion and reduce extracellular matrix remodeling. Taxifolin can significantly improve accelerated aortic aging in VSMC-specific Sfrp1 knockout mice. SFRP1 and / or Taxifolin both play an anti-aortic aging role, prevent and / or treat aortic aging related diseases. SFRP1 can be used as a screening target to screen drugs against cell senescence damage.

This invention provides a pharmaceutical composition for the treatment of hereditary contralateral pigmentary disorders, a mouse model for hereditary contralateral pigmentary disorders, and a screening method for compounds for the treatment of hereditary contralateral pigmentary disorders. [Solution] A heterozygous Adar1 knockout mouse that has lost the function of Adar1 (Adenosine Deaminase Acting on RNA1) throughout the body and exhibits a phenotype of mottled skin and / or fur color.

The application discloses a kind of Thy1-SNCA;Clu gene knockout mouse model construction method and application, the model construction method is: Thy1-SNCA mouse is crossed with Clu-KO mouse and F1 generation is obtained, and Thy1-SNCA;Clu- / + mouse is obtained by identification;Thy1-SNCA;Clu- / + mouse is crossed back with Clu-KO mouse and F2 generation is obtained, and the mouse of target genotype, i.e. Thy1-SNCA;Clu- / - mouse is obtained by identification.The Thy1-SNCA;Clu- / - mouse constructed by the application can effectively shorten the time point of the appearance of parkinsonian movement disorder, thereby shortening the experimental period, and providing more model mouse selection for PD research.

The invention belongs to the technical field of biological medicine, and discloses a construction method and application of a podocyte specific S1PR1 gene knockout mouse model. A podocyte specific S1PR1 knockout mouse is constructed through a Cre-LoxP system, and then an FSGS model is constructed through doxorubicin injection. And verifying the model by using urine detection, serum detection, histopathology detection and molecular mechanism detection. Research finds that S1PR1 expression decline causes POLR2A splicing abnormity and expression decline through down-regulation of Xab2, and podocyte senescence and FSGS progress are induced. According to the invention, a related model is constructed for the first time, the regulation effect of the S1PR1-Xab2-POLR2A pathway in podocyte senescence is disclosed, a reliable tool is provided for researching FSGS pathogenesis, the potential of S1PR1 as a therapeutic target is determined, and the S1PR1 can be used for screening drugs for treating FSGS and has important clinical transformation value.

According to the construction method and application of the animal model for conditional knockout of the Inpp5e gene, a mouse animal model for conditional knockout of the Inpp5e gene is established by applying a CRISPR / Cas9 technology, and specific sgRNA is used; the method comprises the following steps of: constructing a pUC19L-Inpp5e CKO Targeting Vector plasmid by using a seamless cloning method, and constructing a pUC19L-Inpp5e The method for constructing the Inpp5e gene conditional knockout mouse model by using the CRISPR / Cas9 system is simple and easy to implement, the mouse model can be used for researching various neurodevelopment-related diseases, and particularly, a good research model is provided for researching pathogenesis and pathogenesis such as genes on metabolic pathways related to the gene and neurodevelopment defects. And a service is provided for further development of medicines for treating the diseases.

The invention provides application of canthaxanthin in preparation of a medicine for treating and / or relieving xerophthalmia, and relates to the technical field of medicine. According to the application, the distribution condition of an eye tissue structure drug target TMEM16A ion channel is researched, a TMEM16A conditional knockout mouse is constructed through a gene knockout means, and through research on the treatment effect of the knockout mouse, it is verified that canthaxanthin can serve as an activator of the TMEM16A ion channel, and the canthaxanthin can be used as an activator of the TMEM16A ion channel. Therefore, the purpose of application in preparation of the medicine for treating and / or relieving xerophthalmia is achieved.

The invention relates to the field of construction of disease animal models, in particular to a construction method and application of a primary hemophagic syndrome animal model. The model is constructed based on combination of PRF1 gene (perforin 1 gene) knockout mice and lymphochoroidal meningitis virus infection, and typical HLH pathological characteristics are successfully induced in the PRF1 gene knockout mice through single intraperitoneal injection of the lymphochoroidal meningitis virus. Comprise weight loss, continuous fever, pancytopenia, serum ferritin and sCD25 level significant increase and bone marrow blood phagocytosis; compared with an existing CpG ODN1826 repeated injection model, the model has higher genetic specificity and clinical correlation, operation is simplified, cost is reduced, and a standardized experimental platform is provided for pharmacological and pathogenesis research of HLH.

The invention discloses a preparation method of an Erbin gene knockout renal fibrosis animal model. The preparation method comprises the following steps: 1) obtaining an animal of which the Erbin gene is specifically knocked out from renal tubular epithelial cells; and 2) providing an effective dose of cis-platinum for the animal with the Erbin gene specifically knocked out of the renal tubular epithelial cells to obtain the renal fibrosis animal model. The cisplatin-induced renal tubular epithelial cell Erbin-KO specific knockout mouse renal fibrosis model constructed by the invention can be used for further researching the action and mechanism of the Erbin gene in chronic nephrosis, is further applied to the development of drugs for treating and / or preventing renal fibrosis, and has important application prospects and clinical significance.

The invention discloses an application of WBSCR16 protein as a target spot in preparation of a medicine for treating diabetes, and provides an application in research and development of related medicines. By constructing a mouse model of Wbscr16 with pancreatic beta cells specifically knocked out, the action mechanism of WBSCR16 in diabetes mellitus attack is deeply researched, and it is found that WBSCR16 participates in regulating the mitochondrial function of the pancreatic beta cells, and then insulin secretion is affected. On the basis, the invention provides a reagent for regulating the expression of the WBSCR16 protein for treating diabetes mellitus. Meanwhile, a construction method of the Wbscr16 pancreatic beta cell specific knockout mouse model is described in detail, and an important tool is provided for related research. Meanwhile, candidate drugs capable of acting on a WBSCR16 target spot are screened out through drug prediction, and a new strategy and a potential intervention direction are provided for treatment of diabetes mellitus.

The present invention pertains to the technical field of biomedicine. Specifically disclosed is use of a substance for inhibiting the activity and / or expression of a CAPG gene or protein in the preparation of a product for treating renal fibrosis caused by kidney injury. First, research has found that CAPG is expressed at low levels in normal kidney tissues, while its expression level is increased in kidney tissues of patients with acute kidney injury, minimal change disease, IgA nephropathy, lupus nephritis, and diabetic kidney disease, indicating that the expression of CAPG may be involved in the occurrence and development of renal fibrosis caused by kidney injury. Then, by means of constructing a CAPG knockout mouse model, it has been further found that knocking out CAPG can effectively alleviate the progression of renal fibrosis; and by means of culturing renal fibroblasts in vitro, it has been found that silencing or knocking down CAPG can inhibit TGF-β-induced activation of renal fibroblasts. The present invention is of great significance for the pathogenesis and treatment of kidney injury or renal fibrosis.

The invention specifically discloses application of RNA binding protein FXR1 as a target spot in preparation of products for preventing and treating metabolism-related fatty liver diseases and improving glucose and lipid metabolism disorders, and relates to the technical field of biological medicines. By constructing a liver specific knockout mouse (HKO) and high-fat and high-sugar diet induction model, the invention reveals that the RNA binding protein FXR1 is a key pathogenic factor for liver fatty degeneration and metabolic disorder for the first time, and discovers that the deletion of the RNA binding protein FXR1 can significantly improve liver lipid deposition, alleviate liver injury and fibrosis, and can be used for treating liver fatty degeneration and metabolic disorder. The invention can improve the abnormal glucose tolerance and insulin resistance of the whole body, and inhibit the RNA binding protein FXR1 as a new strategy for treating liver glycolipid metabolism and metabolism-related fatty liver diseases.

The application belongs to the technical field of animal models, and particularly relates to a construction method and application of a GRIM-19 and IL-33 double-gene parietal cell-specific knockout mouse model. The construction method comprises the following steps: respectively modifying mouse GRIM-19 and IL-33 genes by flox, then crossing the GRIM-19flox / flox mouse with the IL-33flox / flox mouse to obtain F1 generation mice; crossing the F1 generation mice with Atp4b-Cre mice to obtain F2 generation mice; crossing the F2 generation mice with the F1 generation mice to obtain a GRIM-19 and IL-33 double-gene parietal cell-specific knockout mouse model. The model is helpful for exploring the regulation mechanism of the GRIM-19 and IL-33 genes in SPEM occurrence and progression, and is of great significance for the treatment of SPEM and the prevention and treatment of gastric cancer.

The invention belongs to the field of genetic engineering animal models, and particularly relates to a specific Foxp4 gene knockout mouse model and a construction method and application thereof. The method comprises the following steps: by utilizing a Cre-loxP system, obtaining Foxp4-fl / fl from a Foxp4-fl / fl from a Foxp4-fl / + heterozygote mouse and an Alb-Cre mouse through a mating strategy; and the liver-specific Foxp4 knockout mouse model is constructed by using the Foxp4 knockout mouse and an Albcre / + mouse. The invention finds for the first time that the Foxp4 plays an important role in maintaining the steady state function of the liver, the deficiency of the Foxp4 leads to phenotypes such as lipid accumulation in the liver, inflammatory cell infiltration, liver cell function damage, mild fibrosis and abnormal glucose and lipid metabolism, and the phenotypes are highly consistent with early symptoms and molecular characteristics of NAFLD; the Foxp4 gene has important application value in the aspects of researching the effect of the Foxp4 gene in the occurrence and development mechanism of the NAFLD and the maintenance of liver homeostasis, developing NAFLD-related therapeutic drugs and the like.

The invention belongs to the technical field of molecular biology, and discloses application of DDX24 in maintaining the homeostasis of an intestinal vascular barrier. According to the present invention, the low expression of the intestinal microvascular endothelial DDX24 in the inflammatory bowel disease (IBD) is firstly found so as to construct the adult endothelial cell specific DDX24 induced knockout mouse model, the homeostasis of the intestinal vascular barrier (GVB) of the mouse is imbalanced, and the mouse suffers from spontaneous enteritis, such that the intestinal microvascular endothelial cell DDX24 is the important molecule for maintaining the intestinal homeostasis; and mechanism research finds that the intestinal microvascular endothelial cell DDX24 inhibits cell senescence to protect the GVB steady state. The research discloses an important mechanism of DDX24 for protecting GVB homeostasis by inhibiting intestinal microvascular endothelial cell senescence for the first time, and provides a strategy and basis for clinical diagnosis and treatment of IBD vascular barrier.

This invention belongs to the field of biomedical technology and discloses a method for constructing and applying a podocyte-specific S1PR1 gene knockout mouse model. Podocyte-specific S1PR1 knockout mice were constructed using the Cre-LoxP system, and then an FSGS model was established via doxorubicin injection. The model was validated using urine, serum, histopathological, and molecular mechanism detection methods. The study found that decreased S1PR1 expression induces podocyte senescence and FSGS progression by downregulating Xab2, leading to abnormal POLR2A splicing and reduced expression. This invention is the first to construct such a model, revealing the regulatory role of the S1PR1-Xab2-POLR2A pathway in podocyte senescence, providing a reliable tool for studying the pathogenesis of FSGS, clarifying the potential of S1PR1 as a therapeutic target, and enabling drug screening for FSGS treatment. It has significant clinical translational value.

This invention discloses a mouse experimental method for detecting the tissue distribution of exogenous oxytocin, belonging to the field of biotechnology. Specifically, oxytocin is injected into or nasally sprayed into oxytocin knockout mice, and body tissues, including but not limited to brain tissue, mammary gland tissue, blood, heart tissue, uterine tissue, bone tissue, and muscle tissue, are collected at different time points. The tissue distribution and content of oxytocin are then detected using liquid chromatography-mass spectrometry (LC-MS / MS). This experimental method allows for a wider range of experimental samples, not just focusing on brain tissue and blood; it also has higher sensitivity and stronger specificity; and by utilizing oxytocin gene knockout mice, it eliminates the influence of endogenous oxytocin, providing certain assistance for subsequent animal experiments and clinical research on oxytocin.

The present application discloses use of FcgammaRIIB as an IgG transport receptor in mammary gland of mammals, and belongs to the field of biotechnology. FcgammaRIIB knockout mice and FcgammaRIIB knockout pigs are constructed, and the ratio of mammalian serum IgG to milk IgG in the FcgammaRIIB knockout mice is significantly increased, indicating a decrease in relative milk IgG concentration. Although mammalian serum IgG content is significantly increased in the FcgammaRIIB knockout pigs, IgG in colostrum of mammals is nearly undetectable. Therefore, the present application demonstrates for the first time that FcgammaRIIB serves as the receptor mediating transport of mammalian serum IgG across the mammary epithelial barrier into milk, which addresses a long-standing unresolved issue in the field of maternal passive immunity. Furthermore, the present application provides a new strategy for increasing colostrum IgG content, thereby enhancing early immunity and survival rate of newborn livestock.