355 results about "Knockout mouse" patented technology

The present invention relates to a method for screening therapeutic drugs of schizophrenia using an animal model of the disease. More specifically, this invention relates to a screening method based on the phospholipase C β1 (PLCβ1) knockout mouse as an animal model of schizophrenia with all the major symptoms of the human disease. This knockout mouse exhibits symptoms similar to human schizophrenia such as locomotor hyperactivity, impaired prepulse inhibition of the startle response, lack of barbering and nesting behaviors, socially subordinate status, impaired learning, and lack of type II theta rhythm which has been implicated in working memory. Thus, the knockout mouse of the present invention can be useful as an animal for screening therapeutic drugs against schizophrenia.

The invention relates to a method of constructing an Ifit3-eKO1 gene knockout mouse animal model and belongs to the biotechnical field. The method comprises the following steps: S1, determining specific target sites sgRNA1 and sgRNA2 of a to-be-knocked out gene of an Ifit3-eKO1 mouse, and performing in vitro transcription with Cas9 nuclease to mRNA; and S2, micro-injecting active sgRNA and Cas9RNA into an oosperm of the mouse to obtain the Ifit3-eKO1 gene knockout mouse. The method has the advantages that by using the CRISPR / Cas9 gene knockout technology, the Ifit3-eKO1 gene knockout mouse animal model is constructed for the first time, thereby providing a convenient, reliable and economical animal model for researching action of Ifit3 in tumorigenesis and development.

The invention discloses miR-124 gene knockout murine animal models. The murine animal models refer to mice with miR-124-1, miR-124-2 and miR-124-3 genes knocked out. With the adoption of a Crispr-cas9 gene knockout technology, microRNA (micro ribonucleic acid) miR-124 genes with the highest expression quantity in the brains of the mice are knocked out. The obtained mice show obvious nervous system disease states such as reduction of locomotor activity, deterioration of learning and memorizing abilities, increase of soluble amyloid beta protein and the like. Therefore, the simple, reliable and economical animal models can be provided for research of nervous system disease pathology and screening of drugs for treating nervous system diseases.

The invention discloses a construction method for a Sqstm1 whole-genome knockout mouse animal model and application. The mouse animal model is a mouse of which the Sqstm1 gene is knocked out. On the basis of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 gene knockout technology, the Sqstm1 gene knockout mouse model is constructed. The construction method comprises the following steps: S1: designing sgRNA (Ribonucleic Acid) and Cas9 RNA, carrying out in vitro transcription to obtain mRNA, carrying out microinjection on the sgRNA and the Cas9 RNA with activity into amouse oosperm, to obtain the Sqtm1 gene knockout mouse; S2: identifying the Sqtm1 gene knockout mouse animal model. On the basis of the CRISPR / Cas9 gene knockout technology, the Sqstm1 gene knockout mouse animal model is constructed for the first time, and a convenient, reliable and economic animal model is provided for researching a relationship between Sqstm1 and diseases including autophagy, tumors and the like.

A viable global NaV1.7− / − knockout mouse is disclosed, and a breeding colony of global NaV1.7− / − knockout mice. Also disclosed are an isolated mouse gamete that does not encode a functional NaV1.7− / −, produced by the NaV1.7− / − knockout mouse; an isolated NaV1.7− / − mouse cell, or a progeny cell thereof, isolated from the NaV1.7− / − knockout mouse; and a primary cell culture or a secondary cell line and a tissue or organ explant or culture thereof derived from the NaV1.7− / − knockout mouse. Disclosed also are a hybridoma, wherein the hybridoma was originally formed from the fusion of the isolated NaV1.7− / − mouse cell mouse cell and a myeloma cell, and a method of making an antibody. Also disclosed are assays useful for screening prospective NaV1.7 inhibitors and dose ranging a test NaV17 inhibitor compound, which were validated using the NaV1.7− / − knockout mouse.

The invention relates to a method for acquiring a Nogo-B knockout mode mouse based on CRISPR / Cas9 technology and an application thereof, the method for acquiring the Nogo-B knockout mode mouse comprises the following steps of: designing a high-efficiency sgRNA for identifying EGE-ZXH-007 of a specific gene; connecting the sgRNA to a plasmid vector and then carrying out in-vitro transcription to obtain a Ca9 / sgRNA capable of being subject to microinjection; injecting the Ca9 / sgRNA capable of being subject to microinjection into a mouse fertilized egg, and then carrying out genotype detection and identification on the born mouse, and finally obtaining the Nogo-B knock-out mode mouse. According to the invention, the Nogo-B knockout mode mouse is obtained by using the CRISPR / Case9 technology,and compared with the traditional gene knockout technology, the operation is easy and the efficiency is higher, and the homozygous mutation is more easily obtained, thus laying a good foundation for further researching the function of Nogo-B.

The invention relates to a construction method and application of an immunodeficient mouse animal model, belonging to the field of animal gene engineering and genetic modification. Il2rg gene knockout mice can be obtained by using a CRISPR / Cas9 technology; the Il2rg gene knockout mice have an immunodeficient phenotype. The construction method can be widely applied to multiple fields such as humanized animal models as well as stem cell and tumor research.

The present invention provides an immunodeficient mouse (NOG mouse) suitable for engraftment, differentiation and proliferation of heterologous cells, and a method of producing such a mouse. This mouse is obtained by backcrossing a C.B-17-scid mouse with an NOD / Shi mouse, and further backcrossing an interleukin 2-receptor γ-chain gene-knockout mouse with the thus backcrossed mouse. It is usable for producing a human antibody and establishing a stem cell assay system, a tumor model and a virus-infection model.

This invention relates to transgenic non-human animals comprising a disrupted endothelial nitric oxide synthase gene. These animals exhibit abnormal wound-healing properties and hypertension. This invention also relates to methods of using the transgenic animals to screen for compounds having a potential therapeutic utility for vascular endothelial disorders, such as hypertension, cerebral ischemia or stroke, atherosclerosis and wound-healing activities. Moreover, this invention also relates to methods of treating a patient suffering from hypertension and wound-healing abnormalities with the compounds identified using the transgenic animals, and methods of making the transgenic animals. A method of treating a wound using nitroglycerin is also provided.

This invention provides a therapeutic agent for inhibiting neovascularization, a therapeutic agent for a solid cancer, a therapeutic agent for a disease pathologically caused by neovascularization, and a therapeutic agent for repairing a tissue comprising as the effective ingredient, a substance that potentiates the action of CXCR4.Based on the finding that vascularization is suppressed in CXCR4 knockout mice, it becomes possible to prepare a therapeutic agent for suppressing vascularization, a therapeutic agent for a solid cancer, a therapeutic agent for a disease pathologically caused by neovascularization, each of which comprises as the effective ingredient, a substance that inhibits the action of CXCR4, as well as to prepare a therapeutic agent for repairing a tissue comprising as the effective ingredient, a substance that potentiates the action of CXCR4. Methods for treatment are made possible that use these therapeutic agents.

Disclosed is a transgenic knockout mouse whose genome comprises a homozygous disruption in its endogenous FcRn gene, wherein said homozygous disruption prevents the expression of a functional FcRn protein, resulting in a transgenic knockout mouse in which exogenously administered IgG1 exhibits a substantially shorter half-life, as compared to the half-life of exogenously administered IgG1 in a wild-type mouse. Also disclosed is a transgenic knockout mouse whose genome comprises a homozygous disruption in its endogenous FcRn gene, wherein said homozygous disruption prevents the expression of a functional FcRn protein, resulting in a transgenic knockout mouse which is unable to absorb maternal IgG in the prenatal or neonatal stage of development Methods of using the transgenic knockout mouse, and cells derived therefrom, are also disclosed.

The invention discloses a Cre-LoxP condition-based GLCCI1 gene knockout mouse model constructing kit, and a targeting vector and a constructing method of the above mouse model. A loxP sequence is respectively arranged at two ends of a target sequence of GLCCI1 by using a Cre / loxP principle to obtain a flox mouse, the flox mouse and a mouse with a systematic knockout Cre line are mated to obtain a systematic GLCCI1 gene knockout mouse line, and the obtained GLCCI1 gene knockout mouse is applied to the function researches of the GLCCI1 gene and proteins.

The disclosure concerns a method for resistance of epilepsy by suppressing the function of alpha 1 G protein of T-type calcium channels, use of suppressor of alpha 1 G protein for prevention or treatment for epilepsy, knockout mice resisting epilepsy by disrupting alpha 1 G subunit of T-type calcium channel, and preparation method thereof. In addition, suppressing alpha 1 G protein of T-type calcium channels does not occur epilepsy, and alpha 1 G-deficient mice are useful to study of mechanism related to epilepsy.

The present invention provides an immunodeficient mouse (NOG mouse) suitable for engraftment, differentiation and proliferation of heterologous cells, and a method of producing such a mouse. This mouse is obtained by backcrossing a C.B-17-scid mouse with an NOD / Shi mouse, and further backcrossing an interleukin 2-receptor γ-chain gene-knockout mouse with the thus backcrossed mouse. It is usable for producing a human antibody and establishing a stem cell assay system, a tumor model and a virus-infection model.

By means of a proteomic approach, the markers of hepatocellular carcinoma (HCC) in the liver of a knockout mouse (MAT1A− / −) have been identified for the MAT1A gene (deficient in the synthesis of S-adenosylmethionine). 27 proteins have been detected the expression thereof is altered in, at least, 50% of the analysed tumours. Amongst them, 13 proteins have been validated in biopsies of patients with HCC of different etiology, and 7 of them have been validated in biopsies of patients with liver cirrhosis, a stage prior to the development of HCC, which makes it possible to differentiate between prior stages of the disease and even between different etiologies (viral and alcoholic). Having a panel of markers available may contribute to more accurately defining the alterations associated with the development of HCC and thus facilitate prognosis and diagnosis of this disease.

The disclosure concerns a method for resistance of epilepsy by suppressing the function of alpha 1G protein of T-type calcium channels, use of suppressor of alpha 1G protein for prevention or treatment for epilepsy, knockout mice resisting epilepsy by disrupting alpha 1G subunit of T-type calcium channel, and preparation method thereof. The α1G-knockout transgenic mouse can be used for investigating the relationship between diseases particularly neuropathy or psychopathy and the function of α1G T-type calcium channel via various behavioral tests since α1G subunit is mainly expression in central nervous system (CNS) and pheripheral nervous system (PNS). Further, the α1G-knockout transgenic mouse can be used for screening antileptic agents.

Methods for testing candidate drugs for treatment of age-related macular degeneration are provided. Ccl2-deficient, and Ccr2-deficient mice are used to determine the effect of candidate drugs and treatments on development of age-related macular degeneration. Also provided is a Ccl2-deficient, Ccr2-deficient dual knockout mouse, which is a useful animal model for age-related macular degeneration.

The present invention provides a monoclonal antibody having a pathogenic activity that can induce pemphigus lesion, a peptide specifically recognized by the monoclonal antibody and useful as a therapeutic drug for pemphigus autoimmune disease, etc. As anti-mouse Dsg3 antibody-producing cells are present in the splenocytes of pemphigus vulgaris mouse model constructed by using autoantigen knockout mouse, cell fusion was conducted with the splenocytes of said mouse model and mouse myeloma cells using polyethyleneglycol, hybridomas were constructed, and monoclonal antibodies against Dsg3 were constructed. Among them, a monoclonal antibody having a pathological activity that can induce pemphigus lesions was screened, the base sequence and amino acid sequence in its variable region (heavy chain, light chain) was determined, and the specific epitope part was identified.

The invention relates to a fatty-liver-related liver cancer model building method based on knockout mice. The method is characterized in that APOE- / - and LDLR- / - knockout mice including female mice and male mice are selected, the female mice and the male mice are matched in number, and double knockout mice are obtained; a primer is designed, and the genotypes in mice are identified through a PCR (polymerase chain reaction) system; the PCR (polymerase chain reaction) system comprises an ApoE gene identification PCR conditions and LDLR gene identification PCR conditions; the double knockout mice are pregnant, and suckling mice are obtained; streptozotocin is injected to the suckling mice, the suckling mice begin a high-fat diet after ablactation, and finally the suckling mice surfer from liver cancer; it is identified that the suckling mice begin the high-fat diet after ablactation, fatty liver occurs six weeks later, NASH occurs 8-10 weeks later, dysplastic nodule occurs 13-16 weeks later, and the liver cancer occurs 18-24 weeks later. The occurrence rate of the liver cancer is about 100% 24 weeks later. The time when pure-gene APOE- / - and LDLR- / - knockout mice suffer from NASH and HCC due to the high-fat diet is advanced by 24-30 weeks, and the occurrence rate is high.

An animal model of coronary heart disease has been developed where myocardial infarct can be induced by altering the animal's diet. In all embodiments, this animal model is a result of reduced activity of scavenger receptor class BI (SR-BI) and apolipoprotein E (ApoE). In a preferred embodiment, the model is a result of crossbreeding two transgenic mouse lines: a knockout of SR-BI (SR-BI− / −) and an impaired ApoE expressor (hypoE). The impaired ApoE gene results in only 2-5% expression of ApoE and a reduction in cholesterol homeostasis. Resulting animals are predisposed to hypercholesterolemia but can live longer than a year on a normal low fat diet. Serum plasma levels can be significantly elevated by changing the animal's diet to one containing high levels of fat and cholesterol. Within a month on a high fat, high cholesterol diet, animals develop atherosclerosis and myocardial infarction occurs. Survival depends on the nature of the diet and the conditions of animal husbandry and can typically be around 20-30 days after administration of the modified diet depending on the specific conditions. Housing the animals alone or in groups significantly affects survival of these animals on a high fat diet. Analysis of B- and T-cell deficient SR-BI / ApoE / RAG2 triple knockout mice established that B- and T-lymphocytes do not play a key role in the pathophysiology of the SR-BI ApoE dKO model of human disease. These animal models can be used to study mechanisms and progression of CHD as a function of diet, treatment with drugs to be screened for efficacy or undesirable side effects, and social environmental effects.

The invention discloses a construction method of an Amy1 gene knockout mouse animal model and application. An Amy1 gene knockout mouse model is constructed based on a CRISPR / Cas9 gene knockout technology. The construction method comprises the following steps: step 1, designing sgRNA and Cas9 RNA, performing in-vitro transcription to obtain mRNA, and performing microinjection of the active sgRNA and Cas9 RNA on fertilized eggs of a mouse to obtain an Amy1 gene knockout mouse; and step 2, identifying the Amy1 gene knockout mouse animal model. The method has advantages: the CRISPR / Cas9 gene knockout technology is used and the mouse animal model with the Amy1 gene knocked out is established for the first time; and a convenient and reliable animal model is provided for researching the relationbetween the Amy1 gene and related diseases.

Methods for testing candidate drugs for treatment of age-related macular degeneration are provided. Ccl2-deficient, and Ccr2-deficient mice are used to determine the effect of candidate drugs and treatments on development of age-related macular degeneration. Also provided is a Ccl2-deficient, Ccr2-deficient dual knockout mouse, which is a useful animal model for age-related macular degeneration.

The disclosure concerns a method for resistance of epilepsy by suppressing the function of alpha 1G protein of T-type calcium channels, use of suppressor of alpha 1G protein for prevention or treatment for epilepsy, knockout mice resisting epilepsy by disrupting alpha 1G subunit of T-type calcium channel, and preparation method thereof. The α1G-knockout transgenic mouse can be used for investigating the relationship between diseases particularly neuropathy or psychopathy and the function of α1G T-type calcium channel via various behavioral tests since α1G subunit is mainly expression in central nervous system (CNS) and pheripheral nervous system (PNS). Further, the α1G-knockout transgenic mouse can be used for screening antileptic agents.

The invention relates to a construction method of an AURKA-CKO1-N condition gene knockout mouse model. A recombination targeting carrier is constructed, embryonic stem cell are transfected after linearization, embryonic stem cell with homologous recombination are screened for clone, a donor mice blastula is injected after amplification to produce a chimera mouse; the chimera mouse copulates with an flp mouse to obtain a male F1 generation of mouse; and the male F1 generation of mouse copulates with a mouse whose tissues include specific Cre recombinase, and the AURKA-CKO1-N condition gene knockout mouse model is obtained in the next generation. According to the AURKA-CKO1-N condition gene knockout mouse model, an AURKA gene loses functions in specific tissues and cells, and an ideal modelis provided for research on an action mechanism of Aurora-A in tumorigenesis development.

A viable global NaV1.7− / − knockout mouse is disclosed, and a breeding colony of global NaV1.7− / − knockout mice. Also disclosed are an isolated mouse gamete that does not encode a functional NaV1.7, produced by the NaV1.7− / − knockout mouse; an isolated NaV1.7− / − mouse cell, or a progeny cell thereof, isolated from the NaV1.7− / − knockout mouse; and a primary cell culture or a secondary cell line and a tissue or organ explant or culture thereof derived from the NaV1.7− / − knockout mouse. Disclosed also are a hybridoma, wherein the hybridoma was originally formed from the fusion of the isolated NaV1.7− / − mouse cell mouse cell and a myeloma cell, and a method of making an antibody. Also disclosed are assays useful for screening prospective NaV1.7 inhibitors and dose ranging a test NaV1.7 inhibitor compound, which were validated using the NaV1.7− / − knockout mouse.

The invention discloses novel application of a human YTHDF1 (Youth-Tech-Health Domain Family Member 1) gene, that is, a human YTHDF1 gene expression amount detection reagent is applied to preparationof lung cancer clinical diagnosis reagents, a human YTHDF1 gene is applied to preparation of lung cancer medicines, compared with normal pulmonary epithelial cells, the YTHDF1 gene is highly expressed in lung cancer cells, RNA (Ribonucleic Acid) designed according to the YTHDF1 gene is capable of interfering slow viruses, inhibiting expression of YTHDF1, remarkably inhibiting proliferation of thelung cancer cells, retarding tumor cell division at a G0 / G1 phase and promoting cell apoptosis, inhibiting formation of nude mouse transplanted tumour, and furthermore the purpose of treating lung cancer can be achieved; YTHDF1 gene knockout mouse tumor formation capability detection shows that cell proliferation is remarkably inhibited, and cell apoptosis is remarkably increased; the results show that the YTHDF1 gene can be used as a target for lung cancer treatment, and wide prospects are provided for development of lung cancer treatment medicines based on YTHDF1 genes in future.

The invention discloses a molecular marker for diagnosing atherosclerosis and an application of the marker, belongs to the technical field of biological engineering and relates to an application of areagent for detecting the expression level of SCRAB1 in blood vessel samples to preparation of products for diagnosing the atherosclerosis, and A sequence of the SCRAB1 is as shown in SEQ: ID: NO: 1.ApoE knockout mice and THP-1 source foam cells serve as research objects, RNA (ribonucleic acid) of tissues or cells is extracted, expression change of the gene SCRAB1 related to AS lipid is observed,cellular level validation is implemented, and a deposition specificity diagnosis method of the AS lipid comprising the SCRAB1 is built. The method is simple and reliable, materials are conveniently taken, sensitivity and specificity of the method are highly consistent with those of imaging diagnosis and gross specimen staining, and the method has high application values in early diagnosis and prognosis evaluation of AS diseases.

The invention provides a construction method of an Ano5 gene knockout mouse model. A CRISPR-Cas9 gene knockout technology is adopted to remove 11th and 12th exons of the Ano5 gene. After Ano5 gene knockout, mice all have clinical symptoms of human GDD patients such as lateral bending of shin bones, hyperplasia of cortical bone, hypertrophy of lower jawbone, increasing of alkaline phosphatase in blood, and the like. The established mouse model is used to simulate the human GDD disease, is stable, has stable heredity and similar symptoms of the human GDD disease, can be applied to research on GDD pathogenesis and GDD gene therapy, and is economic, simple and reliable.