122 results about "Mice transgenic" patented technology

Provided in the present invention are a non-human animal expressing a human or chimeric (e.g. humanized) CFB protein and a use method thereof. Further provided in the present invention are a non-human animal genome, a humanized CFB gene, and a cell, tissue, organ or non-human animal containing the non-human animal genome and the humanized CFB gene. The non-human animal obtained by the present application successfully expresses the human or humanized CFB protein, wherein the protein can be cleaved by CFD, bind to C3b, and achieve a similar effect as in a human body. In addition, the non-human animal obtained in the present application has a complete renal function state and normal blood biochemical indexes. Even more rarely, potential diseases known in the prior art which have been observed in some other transgenic mice do not occur in the non-human animal.

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 discloses a construction method and application of a mouse for expressing a heavy-chain antibody IgG1, and relates to the technical field of biology. The mouse capable of expressing the heavy-chain antibody IgG1 is constructed by the method provided by the invention, does not contain an immunoglobulin light-chain gene and a gamma1-CH1 exon, can generate the IgG1 type heavy-chain antibody, and is relatively high in expression quantity and strong in antigen affinity. The invention also provides an application of a mouse expressing the heavy-chain antibody IgG1 in production of a nano antibody. The invention provides a new thought and method for further modifying and transforming transgenic mice expressing more heavy-chain antibodies, and effectively solves the problems of low heavy-chain antibody expression quantity, weak antigen affinity and weak immune response of the existing mice.

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 a construction method and application of a GSDMB intestinal epithelium specific knock-in mouse model. The construction method comprises the following steps: (1) designing gRNA according to an ROSA26 gene, constructing a Donor vector containing a GSDMB gene, introducing the gRNA and the Donor vector and Cas mRNA into a fertilized egg of a receptor mouse, transplanting the fertilized egg and Cas mRNA into a pseudopregnant mouse body for reproduction, and screening out a GSDMB gene knock-in F0-generation mouse; (2) mating and breeding the F0 generation mouse and a wild type mouse, and screening out an F1 heterozygote mouse with GSDMB knocked in; (3) carrying out inbreeding propagation on the F1 heterozygote mouse, and screening out an F2 homozygote mouse in which GSDMB is knocked; (4) the F2 homozygote mouse and the intestinal epithelium specific Vilin-Cre transgenic mouse are subjected to mating breeding, and a heterozygote mouse with GSDMB intestinal epithelium specific knock-in is screened out; and (5) carrying out inbreeding propagation on the heterozygote mouse with the GSDMB intestinal epithelium specific knock-in, and screening out the homozygote mouse with the GSDMB intestinal epithelium specific knock-in.

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 medicine, and particularly relates to application of antifreeze protein in weight losing and blood sugar reducing. The NPAFP transgenic mouse model is successfully constructed by utilizing a genetic engineering technology, and the potential application value of the NPAFP protein in weight loss and blood sugar reduction is verified. Furthermore, based on the model, it is found that the NPAFP protein has remarkable effects in the aspects of reducing fat accumulation and improving the blood glucose level by adjusting brown fat metabolism and the liver lipid droplet content. Based on compound development or protein engineering modification of the NPAFP protein, novel drugs and strategies can be provided for treatment of obesity and diabetes mellitus. Meanwhile, the molecular mechanism of the NPAFP protein is further explored by combining a multi-omics technology, and clinical transformation and application of the NPAFP protein are promoted.

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.

The invention provides a model for monitoring Alzheimer's disease cell cycl-G2 / M period as well as a construction method and application thereof, and belongs to the technical field of biology. According to the invention, an APP / PS1 double transgenic mouse and a Fucci-Green (G2 / M stage) reporter gene mouse are hybridized through a gene recombination technology to obtain an Alzheimer's disease cell cycle G2 / M stage-Fucci-Green reporter gene mouse APP / PS1:: mAG-hGem( 1 / 110), namely a model mouse for monitoring the Alzheimer's disease cell cycle G2 / M stage; the model mouse for monitoring the cell cycle of the Alzheimer's disease can be used for screening Alzheimer's disease treatment drugs, has important significance on mechanism research of the Alzheimer's disease and targeted drug development, and has good practicability.

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 present invention provides affinity matured recombinant monoclonal antibodies (mAbs) and fragments that bind specifically to an HLA-E-peptide complex, including HLA-E-VL9 complexes, and. regulate the cytotoxicity effector cell function of NK and / or CD8+ T-cells positive for cell-surface expression of NKG2A (“NKG2A+”). Herein, monoclonal antibodies were recombinant.lv derived from isolated functional HLA-E-VL9-specific mAbs from HLA-E-VL9 peptide-immunized HLA-B transgenic mice and from the naive human B cell repertoire. Such antibodies are capable of regulating effector cell cytotoxicity' and can preferentially recognize HLA-E-VL9 peptide complexes expressed on the surface of tumor cells. The monoclonal antibodies were subject to one or more rounds of affinity 7 maturation. The invention provides methods for using affinity matured HLA-E-VL9 mAbs to modulate NK and / or CD8+T-cell function as part of immunotherapeutic strategies.

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 method for constructing a transgenic mouse model of a lung tissue specific expression SARS-Cov2 structural protein, which comprises the following steps: inserting an LSL element and gene sequences of SARS-Cov2 structural proteins S, E, M and N into a mouse gene by using a CRISPR / Cas9 technology, mating a positive mouse inserted with LSL-SARSSEMN with a lung type II alveolar epithelial cell specific Cre tool mouse, and constructing a transgenic mouse model of the lung tissue specific expression SARS-Cov2 structural protein. And LSL-SARSSEMN heterozygosis and Cre positive offspring are retained, and mutual breeding is carried out. Detection and evaluation show that the LSL element is successfully cut off by Cre enzyme, and a corresponding mouse shows serious lung injury along with the enhancement of gene expression of each SARS-Cov2 structural protein. According to the invention, the SARS-Cov2 structural protein is specifically expressed in the lung tissue of the mouse, so that the lung injury after SARS-Cov2 infection can be accurately simulated.

The present disclosure provides a transgenic mouse as well as associated methods thereof for use as a mouse model. The transgenic mouse described herein comprises a humanized mouse telomerase (hmTert) gene contained in a background murine germline that provides shortened telomeres that can replicate humans and various human disease states.

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 discloses application of SCTR as an action target in preparation of a medicine for treating pulmonary fibrosis, and belongs to the field of biological medicine. According to the invention, the secretin receptor SCTR is used as an action target for the first time and is applied to preparation of the medicine for treating pulmonary fibrosis. By studying the pathological characteristics of a targeted macrophage Sctr gene knockout transgenic mouse after bleomycin induces idiopathic pulmonary fibrosis, it is found that targeted macrophage Sctr gene knockout can relieve epithelial cell senescence of an idiopathic fibrosis model, improve lung functions and antagonize pulmonary fibrosis progress, and the targeted macrophage Sctr gene knockout has a remarkable effect. The invention provides a new target spot SCTR for the treatment of pulmonary fibrosis, also provides a new thought for searching an action target spot of pulmonary fibrosis and medicine research, and has a wide medical application prospect.

The present invention belongs to the technical field of cell line construction, and particularly relates to a STK11 flox / flox LTF Cre / ‑ Transgenic mouse endometrial tumor cell line and its application. flox / flox LTF Cre / ‑ The deposit number of the transgenic mouse endometrial tumor cell line is CCTCC NO: C2022278. The cell line provided by the present invention has good wall adhesion performance, stable cell characteristics, can be stably passaged for multiple times, has uniform morphological characteristics during passage, can maintain typical characteristics of epithelial cells during passage, has clinical biological characteristics close to tumors, has strong proliferation activity and tumor formation ability, has a high tumor formation rate, is suitable for establishing an endometrial tumor animal model, and provides new ideal experimental materials that are closer to clinical tumor biological characteristics for studying the occurrence and development molecular mechanism of endometrial cancer and screening anti-endometrial cancer drugs.

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.