127 results about "Knockout mouse" patented technology

The invention discloses an application of an HERP gene knockout mouse model in obesity and metabolic disorder diseases. The invention finds that compared with a wild type high-fat group, HERP gene knockout can cause phenotypes such as early obesity, more serious obesity degree, increase of fat weight ratio, aggravation of adipose tissue inflammation, dyslipidemia and glucose metabolism disorder of mice during high-fat feeding. And the HERP gene is knocked out, so that the adipocyte differentiation is increased. The HERP gene knockout mouse is more sensitive to nutrition, obesity can occur more quickly during high-fat feeding, symptoms such as fat amplification, fat inflammation and glucose and lipid metabolism disorder are more serious, and the HERP gene knockout mouse can be used for molecular mechanism research and drug screening of obesity, fat tissue and glucose and lipid metabolism disorder. It is further found that cilostazol can significantly relieve mouse obesity and lipid metabolism disorder caused by HERP gene defects, but has no obvious weight losing effect on normal genotype mice, and a new strategy is provided for precise treatment.

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 field of animal model construction, and particularly relates to application of an NFATC2IP gene knockout mouse model. Experiments prove that after the NFATC2IP gene is knocked out, the mouse has the characteristic changes of ovarian weight reduction, volume reduction, estrus cycle disorder, fertility reduction, AMH reduction, FSH increase and the like. It is indicated that the NFATC2IP gene is a key factor for maintaining ovarian reserve and endocrine balance, and an NFATC2IP gene knockout mouse can be used for revealing the occurrence mechanism of ovarian dysfunction and can also be used for screening of novel ovarian protection drugs, research of a hormone regulation mechanism and discovery and verification of biomarkers.

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 invention provides application of OLA1 as a target spot in preparation of a medicine for inhibiting myocardial fibrosis, relates to the technical field of biomedicine, and aims at solving the problems that in the prior art, the central muscle fibrillation pathogenesis is not completely clear and the treatment means is deficient by constructing a myocardial cell specific OLA1 gene knockout mouse model. And it is proved that mouse myocardial fibrosis can be significantly inhibited by inhibiting OLA1 expression. On the basis, the invention proposes that the OLA1 inhibitor (including small molecule compounds, siRNA and the like) is used for preparing the medicine for treating myocardial fibrosis, and a new treatment strategy is provided for clinic. According to the application, the direct association between the OLA1 gene and the pathological process of myocardial fibrosis is disclosed for the first time, a gene knockout experiment proves that the myocardial fibrosis can be obviously improved by inhibiting the OLA1, and a new direction is provided for developing targeted drugs.

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 invention discloses an application of cGAS as a target in prevention / treatment of Leigh syndrome. The method comprises the following steps: constructing NDufs4; a cGAS DKO gene knockout mouse model is researched, and the result shows that by knocking out cGAS, the glial hyperplasia phenomenon weakening of the mouse with the Like's syndrome can be improved, and neuroinflammation is inhibited. The invention provides a new potential drug target and a treatment strategy for treating the Like's syndrome, and lays a foundation for researching and developing a new drug for treating the Like's syndrome.

The invention discloses a construction method of a spontaneous lupus adipositis animal model, which comprises the following steps: constructing keratinocytes with genotypes of PPAR [gamma] flox / flox- / -Krt5-CreERT < 2 + > / -Adipoq-Cre < + > / -and PPAR [gamma] flox / flox- / -Krt5-CreERT < 2 + > / -Adipoq-CreERT < 2 + > / -and fat cells into a conditional PPAR [gamma]-active receptor gamma gene knockout mouse, so as to construct the animal model with the spontaneous lupus adipositis animal model. Then knocking out PPAR gamma genes in keratinocytes and adipocytes by using a gene knockout activator, so that the mice spontaneously have the phenotypes of lupus septicaemia, including local skin unhairing, lipoatrophy, persistent skin lesion, proteinuria and serum autoantibodies; histological examination finds that lobular adipositis, glomerular volume increase, mesangial cell hyperplasia, inflammatory cell infiltration, glomerular IgG deposition and the like are mainly caused by dermal immune cell infiltration and subcutaneous fat layer lymphocyte infiltration, clinical symptoms of patients with lupus adipositis are simulated, and an important tool is provided for related research of lupus adipositis.

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 present application relates to a kind of construction method of atherosclerosis humanized mouse model, belong to genetic engineering and genetic modification technical field.The present application is first in ApoE and Ldlr double gene knockout mouse background and obtains NOD-4G gene knockout mouse by knockouting Prkdc and Il2rg gene in mouse, humanized atherosclerosis model is constructed by injecting human umbilical cord blood CD133+stem cell and high-fat feed feeding.Through to atherosclerosis degree analysis, it is found that the aorta and aortic root of the humanized atherosclerosis model constructed have a large number of plaque formation, and contain human CD45 + Leukocyte.This atherosclerosis humanized mouse model provides better animal genetic model for the analysis of the role of human immune cells in the occurrence and development of atherosclerosis disease, and has important significance for the research of human disease and the development of new therapy.

This invention discloses a Fam205a1 Methods for constructing and applying gene knockout mouse models. The construction method utilizes CRISPR / Cas9 gene editing technology, targeting mice. Fam205a1 Gene-designed specific sgRNAs for knockout Fam205a1 The regions of exons 1 through 4 of the gene are obtained. Fam205a1 Gene knockout mice. Male mice in this model exhibit infertility, specifically characterized by impaired spermatogenesis, reduced sperm motility, abnormal sperm morphology (particularly head and acrosome defects), and spermatogenic cell apoptosis. However, the weight and appearance of their testes and epididymis show no significant difference from the wild type. The mouse model constructed in this invention has stable genetic traits and a well-defined phenotype, making it an ideal animal model for studying the mechanisms of idiopathic male infertility, screening drugs for treating male infertility, or developing male contraceptives.

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 relates to an application of Fgl2 gene knockout in antagonism of new coronavirus. In the technical scheme provided by the invention, a new crown infected mouse model is established through two SARS-CoV-2 mouse adaptive strains, namely, an MA17 strain and an MASCp36 strain, and it is verified that virus infection can promote Fgl2 expression up-regulation and fibrinogen deposition in mouse lung tissues; in addition, an SARS-CoV-2 mouse adaptive strain MA17 is utilized to infect an Fgl2 gene knockout mouse to prove that the Fgl2 gene knockout can antagonize lung inflammation infiltration, tissue damage, thromboembolism, fibrinogen deposition and fibrosis formation caused by virus infection, so that the occurrence of severe case and death of the mouse is reduced.

The application belongs to the technical field of biotechnology, and discloses application of CRELD2 in preparation of a drug for treating acute pancreatitis. The application constructs Creld2 gene knockout mice and pancreas acinar cell specific Creld2 overexpression mice, and uses caerulein to induce an acute pancreatitis model, so as to clarify the functional role of the Creld2 gene in the caerulein-induced acute pancreatitis, and to prove that Creld2 gene knockout can aggravate pancreas injury, and pancreas acinar cell specific overexpression of Creld2 can reduce pancreas injury. Through the two-way evidence of gene knockout aggravating pancreas injury and gene overexpression reducing pancreas injury, it is firstly proved that CRELD2 has a protective effect on the caerulein-induced acute pancreatitis, and a new strategy for intervention of acute pancreatitis based on CRELD2 is provided.

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 discloses a ribonucleoprotein complex and a construction method of a hand / foot division deformity IV-type animal model, and belongs to the technical field of genetic engineering. According to the mouse model, exons 7 and 8 of the mouse genome Trp63 gene are knocked out, then a sequence is introduced, the 956th basic group is mutated from a G basic group to an A basic group, and the mouse model with the hand / foot division deformity IV is constructed, compared with an existing Trp63 gene knockout mouse model, the mouse model can better simulate the disease, and a foundation is laid for researching the pathogenic mechanism of the disease. A new method is provided for research of limb dysplasia diseases and screening of treatment drugs, and the method has a good application prospect.

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 application discloses a kind of liver disease treatment gene target and drug.The liver function and liver tissue of Ndufa7 gene knockout mouse are detected and analyzed by the present application, and it is found that knockout Ndufa7 gene will cause abnormal liver function of mouse, liver tissue is damaged, and it is found that the expression of Ndufa7 gene can be induced by real-time quantitative fluorescence PCR to induce the occurrence of non-alcoholic fatty liver disease, and Ndufa7 gene can be used as liver disease treatment gene target to develop liver disease treatment drug.In addition, the present application finds that the survival rate of cell under the stimulation of oleic acid can be improved by overexpressing Ndufa7 gene in cell, and oleic acid is a common reagent for inducing non-alcoholic fatty liver cell model, i.e.overexpression Ndufa7 gene can prevent or improve liver disease, and can be used as liver disease treatment drug.The present application fully proves that Ndufa7 is involved in the occurrence of liver disease, and is beneficial to the development of drug for preventing, improving or treating liver disease.