71 results about "Mice transgenic" patented technology

The invention discloses a construction method and application of a mouse model for specifically tracing heart valve cells, and belongs to the technical field of animal model construction. The model construction method comprises the following steps: (1) constructing a Wid1-Cre hybrid mouse before inserting a Cre recombinase coding gene into a termination codon of a mouse Wid1 gene; and (2) hybridizing the Wid1-Cre hybrid mouse with the Rosa26-tdTomato homozygous report mouse, so as to obtain the double transgenic mouse which simultaneously carries the Wid1-Cre gene and the Rosa26-tdTomato gene. According to the invention, on the basis of the expression characteristic that Wif1 is only limited to a valve area in the heart, Wif1-Cre is constructed; according to the Rosa26-tdTomato mouse model, specific tracing of heart valve cells is achieved, and an important experimental tool is provided for mechanism research of heart valve diseases and development of therapeutic drugs.

This invention discloses a method for constructing a biological model of tertiary lymphoid structures, which can rapidly and efficiently induce the generation of in situ TLS, without relying on specific tissues or organs, and without the need for complex and expensive preparation processes such as cell editing and transgenic mice. It can generate stable, mature, and long-lasting TLS structures within 2 to 3 weeks, and has the characteristics of short cycle, low cost, simple operation, reproducibility, safety and effectiveness.

The invention belongs to the technical field of biological genetic engineering, and particularly relates to a mouse model construction scheme capable of reproducing a key phenotype of hemophagocytic syndrome, which comprises the following steps: inserting a loxP-STOP-loxP-LMP1 conditional expression cassette into a Rosa26 safe port site in a fixed point manner to obtain an Lmp1cKI mouse strain; and hybridizing the mouse with an Ncr1-Cre transgenic mouse of NK cell specific expression Cre recombinase. In a double transgenic offspring (Lmp1Ncr1-Cre), STOP fragments are cut off by Cre enzyme, and continuous high expression of EB virus LMP1 protein in NK cells is driven. The model does not need chemical induction or immune intervention, and typical hemophagocytic syndrome phenotypes, namely serum IL-6, TNF-alpha and IFN-gamma, spontaneously appear within 7-9 days after young rats are born; the body weight is reduced and the survival rate is obviously reduced; the proportion of lymphocytes is sharply reduced; and NK cells are depleted. The invention provides a stable and reliable tool for hemophagocytic syndrome pathogenesis research, drug screening and treatment evaluation.

The application relates to a construction method of a Mettl3 transgenic mouse model with specific knockout, enriches the understanding of the function and action mechanism of Mettl3 in mature pancreatic islet beta cells, and provides a new thought and target for the pathogenesis and treatment of diabetes.

The present disclosure provides transgenic mice for use as mouse models, and methods related thereto. The transgenic mice described herein comprise a humanized mouse telomerase (hmTert) gene included in a background mouse germline that provides shortened telomeres capable of reproducing humans and various human disease states.

The invention belongs to the technical field of GPCR drug research and development, and discloses a ratio type cAMP fluorescent probe RaCamp and a construction method of a transgenic mouse, the probe RaCamp is formed by series fusion of G-Flamp2 fluorescent protein and mCherry fluorescent protein through a flexible linker, and the preparation method of the probe RaCamp comprises the following steps: S1, obtaining an mCherry fragment; s2, a G-Flamp2 linear carrier is obtained; s3, homologous recombination; s4, converting and screening; the construction method of the transgenic mouse comprises the following steps: T1, designing a carrier; t2, microinjection of fertilized eggs; t3, embryo transplantation and reproduction; the invention has the beneficial effects that mCherry is added as an internal reference protein on the premise of keeping the sensitivity of the original G-Flamp2 probe, and the selected connecting peptide (GGGGS) 2 enables the expression quantity of the two proteins in cells to be accurately controlled to be 1: 1, so that the problem of background interference in cell observation of in-vivo imaging is well solved. And the sequence is inserted into a Rosa26-LSL box, so that the construction of a Rosa26-LSL-RaCamp mouse is realized.

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 belongs to the technical field of proteomics detection, and particularly relates to a method for detecting mouse liver neomembrane proteome based on an orthogonal translation system and a mouse strain containing an SORT-KASM module. Comprising the following steps: (S.1) constructing a conditionally expressed SORT-KASM transgenic mouse strain; (S.2) activating the expression of the SORT-KAS M by a tissue specific Cre system; (S.3) giving unnatural amino acid ingestion to the mouse; and (S.4) carrying out biotin labeling coupling on non-natural amino acids in the proteome through an azide-alkyne click reaction. According to the detection method, the space and time high resolution of the liver tissue is achieved, the membrane protein has the high-coverage marking capacity, the low-abundance newborn protein has the high-sensitivity marking capacity, good stability is achieved, and the membrane protein participates in the important physiological processes such as substance transfer, energy metabolism and steady state maintaining; the innovation of the detection method provides a new thought for researching the liver cell membrane proteome.

The invention discloses a p16-OSKM transgenic mouse model and a reprogramming senescence cell construction method and application, and belongs to the technical field of cell biology and regenerative medicine. According to the method, a p16-OSKM hybrid mouse capable of specifically inducing an OSKM factor in a p16 high expression cell is obtained on the basis of hybridization of a p16-Ink4a-CreERT2 transgenic mouse, a Rosa26-CAG-LSL-rtTA3-IRES-mKate2 transgenic mouse and a Col1a1-TRE-OSKM-IRES-mCherry transgenic mouse. The selective activation of the OSKM factor in senescent cells with high expression of p16 can be realized by adding doxycycline into primary embryo fibroblasts separated from the mouse under an in-vitro culture condition, so that the senescent cells enter a cell cycle again and recover division and multiplication capacities. The technology provides an accurate model platform for basic research, and can be used for aging mechanism exploration, cell fate regulation and control, chronic disease treatment, development and screening of anti-aging drugs and the like.

The invention belongs to the technical field of gene engineering, and relates to a construction method and verification of a fixed-point knock-in mouse model expressed by fluorescent tracing IFN-gamma protein. According to the invention, a luciferase fluorescent gene is inserted in front of a 3 'UTR (Untranslated Region) of an IFN-gamma gene in a homologous recombination manner by utilizing a CRISPR / Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR associated 9) technology, and connection is carried out through 2A oligopeptide, so that an IFN-gamma-2A-luciferase (IFNG-Luc for short) fluorescent tracing transgenic mouse model is constructed; the model can be used for realizing the co-expression of IFN-gamma (interferon-gamma) and luciferase protein. By utilizing the model, the expression position of the IFN-gamma protein can be marked through the luciferase fluorescence expression, and the expression intensity of the IFN-gamma protein can be reflected through the luciferase fluorescence intensity, so that the effect of the IFN-gamma in specific physiological and pathological processes can be researched.

The present invention provides a basis for using soluble fiber, particularly compositions comprising Agave fructans, to prevent or treat the accumulation of lipids, particularly ceramides in the liver and brain and lipofuscin in the liver, improving the overall integrity of both organs, with treatment being feasible to prevent fatty liver disease and also prevent Alzheimer's disease (AD). In the present invention, we analyze alterations in lipid metabolism in the liver and ceramide production in the brain of transgenic mice (APP / PS1, TG) for AD. Our results indicate that microvesicular lipid accumulation within hepatocytes is increased in TG mice compared to the control group. Furthermore, in the brains of TG mice, a greater accumulation of ceramides is observed in the somatosensory cortex and entorhinal cortex; in the CA1 region of the hippocampus and in the motor cortex, no significant increase was observed when compared to the control group.

This invention belongs to the field of tumor biology and drug screening technology, specifically disclosing a human EGFR mutation-driven mouse primary lung cancer cell line, its construction method, and its applications. The cell line, ZST-1, is a human EGFR (L858R / T790M) mutation-driven lung cancer cell line derived from mouse primary lung cancer. It is stable, capable of subcutaneous tumor formation in C57BL / 6 mice, and simultaneously expresses Luciferase and tdTomato reporter genes. Its construction method includes obtaining transgenic mice, virus-induced tumor formation, continuous in vivo passage in nude mice, and in vitro culture and screening steps. This cell line can be applied to in vitro screening and efficacy evaluation of human EGFR mutation-targeting drugs, research on EGFR-TKI resistance mechanisms, tumor bioluminescence imaging and fluorescence tracing, and in vivo tumorigenesis and efficacy experiments in an immune-intact C57BL / 6 background.

The invention provides a construction method of an atopic dermatitis animal model, and the construction method comprises the step of carrying out sensitization treatment on a TSLP transgenic mouse by adopting local skin stimulation and a mixed sensitization source so as to construct the atopic dermatitis animal model. In the technical scheme, the TSLP transgenic mouse is induced and sensitized by adopting local skin stimulation and a mixed sensitization source, and the skin of the mouse specifically expresses the TSLP, so that the skin inflammation can quickly appear and is similar to the human AD symptom. Compared with a common mouse-induced allergy animal model, the method has the advantages of high sensitization efficiency, obvious sensitization effect and the like, can be applied to experimental research on AD diseases, and provides a research model for clinical research on AD diseases.

The invention discloses a non-human animal integrated with a double-derived VH subclass fusion gene as well as a construction method and application of the non-human animal. The method comprises the following steps: firstly, constructing a gene construct containing human IGHV1-69 and IGHV3-23 fragments, a specific spacer region, a functional element and a rat heavy chain 3'enhancer, and microinjecting the gene construct into a C57BL / 6J fertilized egg with an endogenous heavy chain and a kappa light chain knocked out to obtain a transgenic mouse; through identification, after a mouse endogenous antibody is inactivated, a human gene is stably integrated, a human IgM antibody is expressed and H5N1 HA immunization is carried out, the use frequency of IGHV1-69 is remarkably increased, the mouse produces a high-titer specific antibody, and serum has neutralizing activity. The mouse can generate a target bnAbs aiming at the HA stem and the RBS in parallel, so that the antibody screening efficiency is improved, and an in-vivo screening and evaluation platform for simulating human body fluid response is provided for research and development of influenza antibodies and vaccines.

The invention discloses a ganoderma lucidum preparation for treating Parkinson's disease and application thereof, relates to a medicine, and provides the ganoderma lucidum preparation for treating Parkinson's disease, the ganoderma lucidum preparation comprises ganoderma lucidum hawthorn spore powder and ganoderma lucidum spore oil, the weight ratio of the ganoderma lucidum hawthorn spore powder to the ganoderma lucidum spore oil is (1-2): (1-2), and the weight ratio of the ganoderma lucidum hawthorn spore powder to the ganoderma lucidum spore oil is (1-2): (1-2). In the anti-senile dementia process, it is known that ganoderma lucidum has the protection effect on nerve cell mitochondria, nerve cell axons, nerve cell short processes and the like of a central nervous system, and has the effect of promoting growth of nerve cells of the central nervous system, and the effect is opposite to the pathological development of Parkinson's disease.

The present invention provides non-human animals expressing human CFB protein or chimeric (e.g., humanized) CFB protein and methods for using the same. The present invention further provides a non-human animal genome, a humanized CFB gene, and cells, tissues, organs, or non-human animals containing the non-human animal genome and the humanized CFB gene. The non-human animals obtained in this application successfully express human CFB protein or humanized CFB protein, which can be cleaved by CFD, bind to C3b, and exert similar effects in the human body. Furthermore, the non-human animals obtained in this application have complete renal function, normal blood biochemical indicators, and, more remarkably, do not exhibit the potential diseases observed in other transgenic mice known in this art.

Disclosed is a use of artemether in preventing and treating Alzheimer's disease. Artemether is a safe and effective compound for treatment against malaria. Artemether activates an AMPK signaling pathway in the brain of a transgenic mouse model of Alzheimer's disease, improves learning and memory abilities in the mouse model, reduces neuron apoptosis in the brain, improves function of choline acetyltransferase, inhibits activity of glial cells, and reduces amyloid plaque deposition, neurofibrillary tangle, and neuron loss, thereby providing an experimental basis for clinical research on the treatment of Alzheimer's disease.

The invention discloses application of HLA-DP in neuromyelitis optica diagnosis and recurrence risk assessment, HLA-DPB1 gene polymorphism affects NMOSD susceptibility and disease state, existence of HLA-DPB1 * 0501 allelotype changes expression level of HLA-DPB1 molecules on APC of peripheral blood of a patient, abnormal activation of T cells is caused, blood brain / retina barrier is damaged, and the neuromyelitis optica is diagnosed. Humoral immunity and cellular immunity in the central nervous system are abnormally activated, resulting in the generation of NMOSD focuses and the onset of NMOSD. According to the application disclosed by the invention, an NMOSD mouse model is constructed by using an HLA-DPB1 transgenic mouse to verify that the HLA-DPB1 * 05: 01 allelotype can induce generation of an NMOSD focus and explore a downstream regulatory pathway by changing the expression level of HLA-DP on the surface of APC in peripheral blood, so that a thought is provided for exploring the pathogenesis of the NMOSD and searching a new therapeutic target.

The invention discloses an application of a NanoLuc luciferase double-subunit complementary system in construction of an experimental animal model. The genome of the experimental animal model contains a NanoLuc enzyme large subunit LgBiT encoding gene and stably expresses LgBiT; correspondingly, a tested object used for invading the experimental animal model or connected into the experimental animal model is in fusion connection with NanoLuc luciferase small subunit HiBiT polypeptide serving as a label or a NanoLuc luciferase small subunit HiBiT coding gene serving as a reporter gene, and LgBiT and HiBiT are specifically combined in the experimental animal model and recombined to form the NanoLuc enzyme with catalytic activity. A strong bioluminescence signal is generated under the action of a furazine substrate, so that high-sensitivity, non-invasive and real-time dynamic imaging of in-vivo distribution, loading capacity and diffusion path of a detected object is realized. The invention also discloses a gene editing system for constructing the LgBiT transgenic mouse.

The invention discloses a method for tracing glioma infiltrated microglial cells, and relates to the technical field of biological cell detection. The invention discloses a method for tracing glioma infiltrated microglial cells. The method comprises the following steps: hybridizing to obtain a transgenic mouse for tracing the microglial cells, inducing the microglial cells to express fluorescent protein, constructing an in-situ glioma model, collecting the brain of the mouse, slicing and dyeing. A mouse used in the invention is a transgenic mouse which traces TMEM119 expression, a fluorescent protein tdTomato is labeled on a microglial cell specific marker TMEM119, in a mouse glioma model, the spatial distribution of microglial cells in glioma can be intuitively observed by judging whether tdTomato fluorescence exists or not, and monocyte-derived macrophages are distinguished by co-dyeing with Iba1.

The present invention provides a method for constructing a Map3k3I441M mutation knock-in transgenic mouse model and a use. The method comprises: replacing the region from exon 1 to part of intron 2 of a mouse with a mutant gene fragment, wherein the sequence of the mutant gene fragment is as shown in SED ID NO: 1 in the sequence listing; using a BCA clone as a template to amplify a homologous arm-containing mouse genome fragment as a targeting vector; co-injecting Cas9 and gRNA as well as the targeting vector into fertilized eggs; allowing the fertilized eggs to develop to obtain positive F0 generation samples; co-housing the positive F0 generation samples for breeding to obtain F1 generation samples; performing genotyping on the F1 generation samples to screen for Map3k3I441M samples; and mating the Map3k3I441M samples from the F1 generation samples with a Cdh5-CreERT tool sample to produce F2 generation samples. A model constructed by the construction method can accurately reflect the real situation in a mouse.

The invention belongs to the field of bioengineering, and particularly relates to a construction method and application of a transgenic mouse for screening tissue cell membrane protein in vivo. Aiming at the problems of low in-vivo research flux, unstable library, limited tissue applicability and the like of the traditional membrane protein gene, sgRNA library plasmids are constructed by designing sgRNA of a membrane protein target gene and a control gene; three-positive-mouse containing Cas9, UBC-CreERT2 and sgRNA libraries is obtained through embryo microinjection and hybridization, genotype identification, inducer regulation and control, disease modeling, multi-tissue sample treatment and NGS analysis technical systems are established in a matched mode, and membrane protein gene in-vivo high-throughput screening is achieved. The model can guarantee library stability and screening reliability, covers multi-tissue research scenes, links in-vitro screening and in-vivo verification and is suitable for membrane protein function analysis and disease target mining, the technical process can be popularized to other gene families, and systematic biological research is promoted.

The present disclosure provides immunodeficient NOD.Cg-Prkdc scid Il2rg tm1Wjl / SzJ (NSG TM ) mouse models comprising an inactivated mouse Flt3 allele, a nucleic acid encoding human interleukin 3 (IL3), a nucleic acid encoding human granulocyte / macrophage stimulating factor (GM-CSF), a nucleic acid encoding human stem cell factor (SCF), and an HLA-A2 / H2-D / B2M transgene encoding (i) human B2 microglobulin (B2M) covalently linked to the MHC class I, alpha 1 and alpha 2 binding domains of the human HLA-A2.1 gene and (ii) the alpha 3 cytoplasmic and transmembrane domains of murine H2-Db.

The invention discloses a construction method and application of a sperm-specific hOdf2-GFP transgenic mouse model, and relates to the technical field of reproductive biology and transgenic animal model construction. According to the construction method disclosed by the invention, based on a piggyBac transposon expression vector system, a DNA (Deoxyribose Nucleic Acid) sequence containing a coding human source Odf2 is effectively connected with a Protamine1 promoter. The transgenic positive mouse constructed by the method can specifically express human Odf2, has a dynamic change from sperm to zygote to eight cell stages, realizes early continuous visual tracking of embryos, has genetic stability, keeps expression and phenotype consistency across generations, and has a good application prospect. The method can be applied to the fields of environmental poison evaluation, asthenospermia mechanisms, drug screening and the like.

The invention belongs to the technical field of gene editing, and particularly relates to a construction method of a transgenic mouse with a tissue-specific precancerous marker. A fertilized ovum is constructed through a gene editing technology, a mutant p53 protein and a fluorescent protein are fused, a loxp-Stop-loxp sequence is added in front of a promoter of a mutant p53 sequence, the fertilized ovum is transplanted into a female mouse, the obtained progeny is an LPL transgenic mouse, and expression inhibition of the mutant p53 protein and the fluorescent protein is achieved; the LPL transgenic mouse is mated with a Cre tool mouse to obtain a transgenic mouse of which the offspring is a tissue-specific precancerous marker, the transgenic mouse expresses mutant p53 protein and fluorescent protein at a specific tissue, early tumor focus development is monitored and traced through a living body fluorescence imaging system, and the LPL transgenic mouse has extremely high application and research values.

The invention relates to the technical field of disease animal model construction methods, and provides a construction method of a rheumatoid arthritis and atherosclerosis disease model, which is characterized in that an hTNF-a transgenic mouse and an APOE transgenic mouse are used for mating and breeding, and the hTNF-a and APOE transgenic mice are screened out for modeling. The modeling method provided by the invention can be close to a model of human rheumatoid arthritis combined with cardiovascular diseases, and modeling is simple and practical.

This invention discloses the application of the small molecule compound Z2114502286 in the preparation of drugs for the prevention and treatment of liver-related diseases. This invention reveals for the first time that Z2114502286 is a THBS2-specific small molecule inhibitor and verifies its feasibility in preventing and treating liver fibrosis by targeting and inhibiting the THBS2 protein. Through virtual screening and molecular docking, this compound can directly bind to the THBS2 protein. In vitro and in vivo studies show that this compound inhibits THBS2-mediated HSC activation in a concentration- and time-dependent manner, inhibits ECM deposition in a mouse model of liver fibrosis, downregulates the expression of fibrosis-related genes, and alleviates liver inflammation and macrophage infiltration. Simultaneously, the targeted efficacy was verified in a transgenic mouse model specifically expressing human THBS2, demonstrating significant clinical translational value. Furthermore, it exhibits bioavailability and safety at effective dosages, showing broad clinical application prospects.

The invention discloses a construction method of a Rosa26-APP-eta-CTF transgenic mouse model and an application of the Rosa26-APP-eta-CTF transgenic mouse model. According to the invention, a CRISPR / Cas9 mediated embryo gene editing technology is utilized, an expression cassette containing a human-derived APP-eta-CTF-mCherry fusion gene is inserted into a mouse Rosa26 site at a fixed point through a homologous recombination repair mechanism, and a transgenic mouse model capable of inducing expression is constructed. The model fills the blank of lack of an animal model for specifically simulating in-vivo accumulation of eta-CTF in the prior art, and provides a key tool for researching the effect of an eta-secretase pathway in the Alzheimer's disease.

Hepcidin is an hyposideremic hormone made primarily by the liver. To address the role of hepcidin made specifically by the intestine in lowering serum iron, the inventors generated transgenic mice overexpressing the peptide specifically in this tissue. These mice exhibit, at one month of age, a severe hyposideremia, along with decreased haematological indices and hair loss. Mechanistically, they showed that intestinal hepcidin made by the transgenic mice had no effect on intestinal ferroportin, but, in contrast, induced a striking down-regulation of Divalent Metal Transporter 1 (DMT1) protein at the apical side of the enterocyte. Intestinal hepcidin can be produced in the apical side suggesting the direct role of apical hepcidin on DMT1. To confirm the therapeutic capacity of hepcidin on regulation of DMT1, the inventors developed probiotics (engineered recombinant lactic acid bacteria), capable of delivering hepcidin directly into the lumen of the intestine. They orally administrated daily these probiotics in hemochromatosis mice model, after 28 days of treatments they observe a decrease of iron overload. Thus, the present invention relates to a method for preventing or treating an iron overload associated disease in a subject in need thereof, comprising administering locally in the gut of the subject hepcidin.

The invention discloses a construction method of a microglial cell conditional knock-down dynactin animal model, which comprises the following steps: selecting a Dctn1LoxP gene knock-in mouse, hybridizing the Dctn1LoxP gene knock-in mouse with a Cx3cr1-Cre transgenic mouse, and carrying out three rounds of breeding to obtain a genotype Dctn1LoxP / LoxP; a mouse of Cx3cr1-Cre; the expression of Cre recombinase is driven by a Cx3cr1 gene promoter, and the 2nd to 4th exons of the Dctn1 gene are cut, so that the knockout of p150Glue and the reduction of the protein levels of other subunits of dynactin are realized. The model is strong in specificity and high in knock-down efficiency, animals show microglial cell morphology abnormity and movement coordination function defects, the model can be used for researching the influence of dynactin on microglial cell neurobiological functions, and a reliable tool is provided for pathogenesis exploration and drug screening of related neurological dysfunction.