285 results about "Knockout animal" patented technology

Disclosed herein are methods for generating recombinant DNA molecules in cells using homologous recombination mediated by recombinases and similar proteins. The methods promote high efficiency homologous recombination in bacterial cells, and in eukaryotic cells such as mammalian cells. The methods are useful for cloning, the generation of transgenic and knockout animals, and gene replacement. The methods are also useful for subcloning large DNA fragments without the need for restriction enzymes. The methods are also useful for repairing single or multiple base mutations to wild type or creating specific mutations in the genome. Also disclosed are bacterial strains and vectors which are useful for high-efficiency homologous recombination.

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 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.

The invention discloses a method for constructing a mouse model for conditional knockout of CCR5 gene of endothelial cell. The method comprises the following steps of firstly, obtaining a CCR5loxp/loxp mouse, and mating with a Tie-2-cre/ERT2 mouse, so as to obtain heterozygous mice with specific knockout of CCR5 gene of the endothelial cell; mutually mating the heterozygous mice, so as to obtain homozygous mice with specific knockout of CCR5 gene out of the endothelial cell. The constructing method disclosed by the invention has the advantages that the mouse with conditional knockout of endothelial cell gene is constructed via the CRISPR/Cas9 system, the mutation is led into the mouse by a cell specificity method, the missing of the CCR5 target gene occurs at the certain tissue organ of the test animal, the controllability of mechanism study is furthest realized, and the disadvantages that different tissues or cells are not distinguished, and target genes in all tissues or cells of themouse body are all removed in the conventional gene knockout technique is overcome.

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.

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.

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.

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.

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.

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.

A viable global Na_V1.7^−/− knockout mouse is disclosed, and a breeding colony of global Na_V1.7^−/− knockout mice. Also disclosed are an isolated mouse gamete that does not encode a functional Na_V1.7, produced by the Na_V1.7^−/− knockout mouse; an isolated Na_V1.7^−/− mouse cell, or a progeny cell thereof, isolated from the Na_V1.7^−/− knockout mouse; and a primary cell culture or a secondary cell line and a tissue or organ explant or culture thereof derived from the Na_V1.7^−/− knockout mouse. Disclosed also are a hybridoma, wherein the hybridoma was originally formed from the fusion of the isolated Na_V1.7^−/− mouse cell mouse cell and a myeloma cell, and a method of making an antibody. Also disclosed are assays useful for screening prospective Na_V1.7 inhibitors and dose ranging a test Na_V1.7 inhibitor compound, which were validated using the Na_V1.7^−/− knockout mouse.

Uteroglobin has been discovered to prevent IgA mediated diseases, such as IgA nephropathy, by preventing the deposition of IgA-Fibronectin immunocomplexes in tissues such as the renal glomeruli. The invention therefore includes methods of treating such diseases by administering therapeutically effective amounts of uteroglobin (and variants or mimetics) to prevent or improve the IgA mediated condition. Transgenic uteroglobin knockout animals, and animals in which uteroglobin-protein expression is reduced by antisense technology, also provide systems for studying IgA mediated diseases, and screening for appropriate treatments.

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.