80 results about "Knockout mouse" patented technology

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.

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 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 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 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 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.

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 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.

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 long-chain non-coding RNA (Ribonucleic Acid) and application thereof. According to the invention, by analyzing and comparing the expression conditions of the long-chain non-coding RNA in a wild type mouse and an ER beta gene knockout mouse, the long-chain non-coding RNA participating in the regulation and control of the osteogenic differentiation of the bone marrow mesenchymal stem cells, namely the long-chain non-coding RNA MSTRG.47495.1 (lncRNA MSTRG.47495.1), is found. Overexpression of the long-chain non-coding RNA in the bone marrow mesenchymal stem cells can effectively promote osteogenic differentiation of the bone marrow mesenchymal stem cells, enhance bone regeneration and promote bone defect repair of rats with skull injury. On the basis, the long-chain non-coding RNA can promote osteogenic differentiation of bone marrow mesenchymal stem cells and enhance bone regeneration, and is used for preparing medicines for treating bone metabolic diseases and bone formation disorder related diseases. In addition, the long-chain non-coding RNA can also be used for construction of bone organs.

The invention relates to the technical field of animal models, and particularly discloses a construction method and application of a mouse model with fat dystrophy complicated with atherosclerosis, and the construction method comprises the following steps: mating MED1flox / flox mice with Adipoq-CreERT2 mice to obtain F1-generation mice; the F1-generation mice and MED1flox / flox mice are subjected to backcross, the mice with the genotype being MED1fl / flAdipoq-Cre < + / -> are screened, tamoxifen is injected to induce fat cell specific Cre recombinase expression, and fat cell specific MED1 gene knockout mice are obtained; and performing caudal vein injection of AAV8-PCSK9 virus and feeding with a high-cholesterol and high-fat feed to obtain the fat dystrophy complicated with atherosclerosis mouse model. According to the method provided by the invention, the mouse model with fat dystrophy complicated with atherosclerosis is successfully constructed and obtained, and collaborative simulation and regulation of phenotypes of two diseases are realized.

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 invention relates to application of a JAML gene in preparation of an abdominal aortic aneurysm resisting medicine, and belongs to the technical field of biological medicine. The nucleotide sequence of the JAML gene is as shown in SEQ ID No.1. The application of the JAML gene in preparation of the abdominal aortic aneurysm resisting medicine comprises the following two aspects: (1) applying the JAML gene as an action target to preparation of the abdominal aortic aneurysm resisting medicine; and (2) applying the JAML gene as an action target to screening of drugs for resisting abdominal aortic aneurysm. According to the invention, a gene knockout mouse of which the smooth muscle specifically knockout JAML is constructed, and then the mouse abdominal aortic aneurysm model is constructed by subcutaneously pumping AngII, so that the morbidity of abdominal aortic aneurysm can be obviously reduced after the JAML gene is knocked out; therefore, a theoretical basis is provided for preparing or screening drugs or preparations for inhibiting smooth muscle cell JAML gene expression as alternative drugs or preparations for resisting abdominal aortic aneurysm.

The application discloses sgRNA for knocking out a TMEM91 gene and application thereof, and relates to the technical field of biological medicines.The sequence of the sgRNA is shown in SEQ ID No.1 or SEQ ID No.2.A gene knockout mouse is constructed according to the TMEM91, and experimental results prove that the sequence can significantly silence the expression of TMEM91.Compared with wild-type mice, the TMEM91 mRNA of the knockout TMEM91 mouse is completely removed, and the TMEM91 gene knockout mouse provides new evidence for the biological function of TMEM91 at the in-vivo level.The application proves for the first time that inhibiting the expression of the TMEM91 gene can enhance kidney injury of the mouse, thereby proving that the TMEM91 is a new target for treating kidney injury, and the application is beneficial to further studying the mechanism of occurrence and development of kidney injury related diseases caused by low expression of the TMEM91.

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 discloses a method for establishing an inflammation model of a Gga3 gene knockout mouse and application, and relates to the technical field of biological medicine, and the key point of the technical scheme is as follows: it is found that Gga3 gene knockout significantly reduces inflammation in the brain of the mouse caused by LPS, and it is prompted that GGA3 plays an important role in regulating and controlling neuroinflammation. The regulation mechanism of GGA3 in microglial cell mediated neuroinflammation can be better illuminated, and the GGA3 has a very important effect on finding drug targets of related neurodegenerative diseases.

The invention discloses an intestinal cancer mouse model based on Par3L gene knockout and a construction method thereof. After sgRNA is designed to knock out a promoter and a first exon of a Par3L gene of a mouse, the progeny of the obtained gene knockout mouse can spontaneously form intestinal cancer along with the increase of age, and the progeny is accompanied with a tissue infiltration phenotype. The intestinal cancer mouse model constructed by the invention can cause abnormal mitosis of intestinal cells and influence the genome stability, a good model system is provided for further research on the occurrence mechanism of intestinal cancer, and the intestinal cancer mouse model has a wide application prospect.

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 application discloses a kind of construction methods of mouse model for studying vascular endothelial senescence, belong to genetic engineering technical field, utilize 6030442E23Rik conditional gene knockout mouse and Tek-CreERT2 mouse (utilize CRISPR gene editing technology, Cre is inserted into mouse Tek gene stop codon, is the tool mouse of endothelial cell conditional knockout, currently, this kind of strain mouse has on market), cross, obtain 6030442E23Rik- / -Tek-CreERT2 mouse.Tamoxifen is dissolved in corn oil later, so that its final concentration is 20 mg / ml.Tamoxifen dose is 120mg / kg mouse weight later, intraperitoneal injection 5 times, once every other day, and the mouse model of endothelial cell 6030442E23Rik specificity knockout can be obtained 7 days after the end of last injection.The mouse model of endothelial cell 6030442E23Rik specificity knockout is an important mouse model for studying vascular endothelial senescence.