112 results about "Knockout animal" patented technology

The invention relates to the technical field of life science, in particular to a method for constructing an atrophic gastritis-based SPEM lesion model, which comprises the following steps: S1, constructing an Slc7a11 conditional knockout mouse strain: carrying out cage hybridization on an Slc7a11 flox / flox mouse and an ATP4b-Cre transgenic mouse, and knocking out a gastric wall cell specific Slc7a11 gene; s2, carrying out genotyping identification; the identification method based on the atrophic gastritis SPEM lesion model and the construction method based on the atrophic gastritis SPEM lesion model comprise the following steps: S1, pathological model verification: taking a mouse gastric mucosa tissue slice, quantitatively analyzing the proportion of an SPEM region by adopting an immunofluorescence double-standard technology, and confirming Slc7a11 knockout induced SPEM pathological characteristics in combination with the existence of SPEM in the gastric mucosa; and S2, long-term pathology evolution monitoring. Through gastric wall cell specific Slc7a11 gene knockout and multiple PCR identification technologies, the initial response SPEM state after gastric mucosa damage is successfully simulated, and the kit is used for researching atrophic gastritis pathological mechanisms and evaluating drug intervention.

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 discloses application of a Foxn3 gene or protein in preparation of a medicine for treating retina ciliary lesion. Experimental results show that Foxn3 is crucial to inhibition of cilia gene expression in non-photosensitive neurons such as bipolar cells and non-long-process cells, neuron functions in retinas of Foxn3 knockout mice are damaged, and deletion of Foxn3 causes ectopic cilia gene expression and abnormal cilia of the non-photosensitive neurons. The invention discloses a promoter directly combined with Foxn3 and inhibiting cilia genes and cilia transcription factors. The promoter comprises Foxj1 and Rfx family members. Foxn3 is used as a key transcription inhibition factor, and a proper cilia structure of the retinal neuron is ensured by preventing non-photosensitive neurons from adopting photosensitive cell-like cilia characteristics. The invention provides a therapeutic target and a drug for retina ciliary lesion.

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 invention relates to the field of biological medicine, in particular to a GDF-15 defect-based modeling method which comprises the following steps: ligating left coronary anterior descending branches of a plurality of GDF-15 knockout mice, and respectively establishing myocardial ischemia reperfusion models based on recombinant GDF-15 and normal saline; the method comprises the following steps: taking a myocardial tissue sample, and treating the sample at a pre-plating temperature and a pre-plating duration to obtain myocardial cells; detecting an AMPK signal channel expressed by the myocardial tissue protein; determining whether the activity of the AMPK signal path is inhibited or not based on the phosphorylation state value, and determining the osmotic pump rate; or, detecting the ROS level of the myocardial cells to obtain a captured image, determining the state of the myocardial cells based on the gray value of the image, and determining the stability of the state of the myocardial cells according to the oxidative stress index of the captured image; carrying out an anoxic experiment to obtain a fluorescence image; and determining the myocardial cell purity based on the fluorescence signal intensity, adjusting the osmotic pump rate, or adjusting the pre-plating time length or the pre-plating temperature, so that the accuracy of GDF-15 defect modeling drug administration is improved.

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 invention provides a method for constructing an organ regeneration genetic compensation effect, and relates to the technical field of biology. Comprising the following steps: S1, designing sgRNA; s2, carrying out in-vitro transcription and purification to obtain a high-purity sgRNA solution; s3, preparing fertilized eggs, preparing sgRNA mixed injection, and performing embryo culture after micro-injection operation to obtain F0-generation animal larvae; s4, performing gene knockout efficiency detection on the F0-generation animal larvae, screening out the F0-generation animal larvae with high knockout efficiency, and breeding and mating the F0-generation animal larvae to obtain F1-generation animals; and S5, verifying and screening the F1-generation animals to obtain homozygous knockout individuals, carrying out amputation treatment on the homozygous knockout individuals, and verifying the genetic compensation effect of organ regeneration. The YAP knockout animal is constructed through a gene editing technology, the effect and mechanism of the YAP knockout animal in limb regeneration are researched, whether YAP deletion affects the regeneration capacity through a genetic compensation effect or not can be revealed, and then a theoretical basis and an experimental basis are provided for future regenerative medicine application.

The present invention belongs to the field of biomedical technology and specifically discloses the use of a substance that inhibits the activity and / or expression of a CAPG gene or protein in preparing a product for treating renal fibrosis caused by renal injury. The present invention first discovered through research that CAPG is expressed at a low level in normal renal tissue, but its expression level is elevated in renal tissue of patients with acute renal injury, minimal change disease, IgA nephropathy, lupus nephritis, and diabetic nephropathy, indicating that CAPG expression may be involved in the occurrence and development of renal fibrosis caused by renal injury. Then, by constructing a CAPG knockout mouse model, it was found that knocking out CAPG can effectively reduce the progression of renal fibrosis. By culturing renal fibroblasts in vitro, it was found that silencing or knocking down CAPG can inhibit TGF-β-induced renal fibroblast activation. The research of the present invention is of great significance for the pathogenesis and treatment of renal injury or renal fibrosis.

The invention discloses application of DPP3 as a target spot in preparation of a traditional Chinese medicine for treating and / or preventing pyemia hypersomnia, and relates to the technical field of biomedicine.In-vivo experiments find that the expression level of DPP3 in serum of a pyemia patient is remarkably and positively correlated with the severity of the disease, and the DPP3 has a significant positive correlation with the severity of the disease. Therefore, the DPP3 can be used as a biomarker of severity of the severity of the severity of the severity of the severity However, whether the elevated DPP3 itself will produce a biological effect has not been clear. Based on this, the invention explores the effect of DPP3 with increased septicopyemia in septicopyemia hypersomnia, and finds that after septicopyemia, the DPP3 content in plasma is increased, a hypersomnia phenomenon occurs, and after septicopyemia is induced by a DPP3 knockout mouse, sleep in a dark period is reduced, and a waking period is increased. In addition, after the DPP3 is intravenously injected into the tail of the mouse, non-rapid eye movement sleep in the dark period is increased, and the waking time is shortened. The invention provides a new molecular target and intervention means for the treatment of pyemia-related hypersomnia, and has important clinical application value.

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 invention discloses application of a biomarker LKB1-PD-1 signal axis as a marker for early diagnosis of obesity and type 2 diabetes mellitus and as a type 2 diabetes mellitus treatment target. The invention also discloses a medicine containing the biomarker, a medicine composition, a detection reagent, a kit, an LKB1-PD-1 signal axis regulator, a mouse model R5 / + Stk11f / f with specific deletion of type-2 intrinsic lymphoid cells (ILC2), a mouse model Id2Cre-ERT2Stk11f / f with ILC2 induced knockout of LKB1 and application of the mouse model. The invention also discloses an application of the biomarker LKB1-PD-1 signal axis in preparation of drugs for diagnosis, prevention and treatment of type 2 diabetes mellitus. The invention has a wide application prospect in the field of diagnosis and treatment of obesity and type 2 diabetes mellitus.

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 present invention discloses a method for constructing a conditional knockout mouse model of the Cpt1a gene and its application. The construction method is to construct a lupus erythematosus mouse model by conditionally knocking out the Cpt1a gene in macrophages. The construction method of the lupus erythematosus mouse model provided by the present invention realizes the knockout of the Cpt1a gene in the bone marrow cell lineage, while the gene is normally expressed in other tissues or cells. It is a simple, efficient and rapid construction method. The obtained lupus erythematosus mouse model can be better applied to the mechanism research of lupus erythematosus, and the preparation and / or screening of drugs for treating lupus erythematosus.

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 present invention relates to the field of genetic engineering technology, and particularly to a method for constructing a mouse model with specific knockout of the Osgep gene in pancreatic islet β cells and its application. The construction method includes the following steps: 1) Determine the knockout region as the exon 2 and exon 3 regions according to the structure of the Osgep gene. The CDS sequence of the Osgep gene transcript is shown in SEQ ID NO.1. Design a gene fragment with flox sites connected at both ends and containing the sequence of the knockout region, and insert this fragment into a plasmid; 2) Design sgRNA according to the upstream sequence and downstream sequence of the knockout region; 3) Mix the sgRNA, Cas9 enzyme and plasmid, and inject them together into the cytoplasm of mouse fertilized eggs to construct edited mouse fertilized eggs and produce F0 generation mice. The present invention discovers through research that Osgep knockout mice can be used for the research of diabetes, and the Osgep gene can play a role in treating diabetic mice.

The present invention belongs to the field of animal genetic engineering and genetic modification. Specifically, it discloses a method for constructing a humanized epidermolysis bullosa DEB mouse model and its application in biomedicine. The method comprises the following steps: preparing genetically engineered mice using mouse embryonic stem cells, and replacing the full-length mouse Col7a1 gene sequence with a human COL7A1 genomic sequence or a mutant sequence thereof; preferably, the human COL7A1 genomic sequence is represented by hg19:chr3:48,635,720-48,603,572; the human COL7A1 genomic mutant sequence is represented by hg19:chr3:48,635,720-48,603,572&c.6527 dup C; the mutant animals constructed by the present invention can survive for 7-10 days, which is significantly longer than the median survival of 2 days reported for Col7a1- / - knockout mice, without drug maintenance, providing a longer window of survival for therapeutic experiments.

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.