96 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.

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.

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 invention belongs to the technical field of cell engineering, and particularly relates to a construction method and application of a mammal rapamycin target protein self-activated mouse model and a mouse embryo fibroblast line. According to the invention, serine (Serine, S) at site 2215 of mTOR protein is mutated into tyrosine (Tyrosine, Y), and a transgenic mouse model with activation mutation of mTORS2215Y, namely a mouse model with self-activation of mammal rapamycin target protein, is constructed. A mouse embryo fibroblast line which can be immortalized is constructed by utilizing the transgenic mouse model, so that the related mechanism research of mTOR self-activation mutation can be carried out at the cellular level, and the screening of related therapeutic drugs can be carried out.

Provided herein, in some aspects, is a NOD.Cg-Prkdcscid Il2rgtm1wjl / SzJ (NOD scid gamma or NSG™) mouse comprising a nucleic acid encoding human FLT3L and an inactivated mouse Flt3 allele, methods of producing the mouse, and methods of using the mouse.

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 construction method and application of an enterovirus 71 type susceptible transgenic mouse model, and belongs to the technical field of biology. The method comprises the following steps: up-regulating or enhancing or improving the content or / and activity of placenta specific protein 8 in a receptor mouse to obtain a mouse overexpressing the placenta specific protein 8, hybridizing the mouse overexpressing the placenta specific protein 8 with a wild mouse to obtain an F1-generation heterozygote mouse, a homozygote mouse obtained after the F1-generation heterozygote mice are hybridized with each other is an enterovirus 71 type susceptible transgenic mouse model. The application has the advantages that not only is the susceptibility of mice to EV71 increased, but also a proper animal model is provided for exploring the important function of Plac8 in the EV71 infection process.

The present invention belongs to the field of biomedical technology, and specifically relates to the application of an antifreeze protein in weight loss and blood glucose reduction. The present invention successfully constructs an NPAFP transgenic mouse model by using genetic engineering technology, and verifies the potential application value of the NPAFP protein in weight loss and blood glucose reduction. Further, based on this model, it is found that the NPAFP protein has a significant effect on reducing fat accumulation and improving blood glucose levels by regulating brown fat metabolism and liver lipid droplet content. The development of compounds or protein engineering modification based on the NPAFP protein of the present invention can provide new drugs and strategies for the treatment of obesity and diabetes. At the same time, further exploring the molecular mechanism of the NPAFP protein by combining multi-omics technologies helps to promote its clinical translation and application.

The invention discloses a GATA4 gene-based MAFLD animal model construction method and application in prevention and treatment drugs, and belongs to the field of mouse model construction, and the GATA4 gene-based MAFLD MASH animal model construction method comprises the following steps: S1, hybridizing a GATA flox transgenic hybrid mouse and a Vil1-Cre transgenic mouse to obtain an F1 generation, and hybridizing the GATA flox transgenic hybrid mouse and a Vil1-Cre transgenic mouse with the genotype of GATA4 flox / +, Vil1-Cre transgenic mouse in the F1 generation to obtain a GATA4 flox / +, Vil1-Cre transgenic mouse to obtain a GATA4 flox / +, Vil1-Cre transgenic mouse; the method comprises the following steps: S1, carrying out selfing on transgenic mice with GATA4 flox / flox and Vil1-Cre to obtain F2-generation mice, and S2, sequentially giving GATA4 intestinal specific knockout mice with genotypes of GATA4 flox / flox and Vil1-Cre in the F2-generation mice with high-fat and high-cholesterol feed (HFD), and inducing to obtain an intestinal GATA4 specific knockout metabolism-related steatohepatitis (MASH) and hepatic fibrosis mouse model. The invention provides a model construction method of MASH and hepatic fibrosis animals in which the intestinal GATA4 gene is specifically knocked out, and application of the intestinal GATA4 serving as a target to preparation of drugs for preventing and treating related diseases such as liver injury and liver diseases caused by the MASH, and the intestinal GATA4 has an important clinical prospect.

The invention discloses a construction method of a metabolism-related fatty liver disease and chronic hepatitis B-related hepatocellular carcinoma model, and belongs to the technical field of disease animal models. The construction method comprises the following steps: S1, linearizing an HBV genome, and injecting the linearized HBV genome into a fertilized egg of a C57BL / 6 mouse to establish an HBV transgenic mouse; and S2, feeding the HBV transgenic mouse in the step S1 with D09100310 feed to obtain the mouse model of metabolism-related fatty liver disease combined with chronic hepatitis B-related hepatocellular carcinoma. According to the construction method of the metabolism-related fatty liver disease combined chronic hepatitis B related hepatocellular carcinoma model, HBV-Tg mice are selected as experimental animals, the D09100310 mouse feed is utilized to construct the metabolism-related fatty liver disease combined chronic hepatitis B related hepatocellular carcinoma model, the stability and repeatability are good, objectiveness and simplicity are achieved, operability is high, and the model is suitable for being used for the clinical application. And other possible interference factors are reduced.

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.