7 results about "Wild type mice" patented technology

The application relates to a construction method of a conditional overexpression ZFP595 transgenic mouse model and application thereof, and relates to the technical field of transgenesis.The construction method comprises the following steps: constructing a homologous recombination vector containing a ZFP595 gene, wherein the vector comprises a 3.3 kb 5' homologous arm, CAG-LSL-Zfp595-HA-IRES-tdTomato-Wpre-pA and a 3.3 kb 3' homologous arm; microinjecting Cas9 mRNA, gRNA and the homologous recombination vector into a zygote of a recipient mouse to obtain F0 generation mice, and performing identification; mating the F0 generation mice identified as positive with wild type mice and backcrossing to obtain a mouse model for conditionally overexpressing ZFP595.The conditional overexpression ZFP595 mouse model constructed by the construction method can be crossed with various Cre mice to realize specific overexpression in different tissues, organs and cells.

The invention belongs to the technical field of biological medicines, and relates to application of Kir4.2 in treatment of renal fibrosis. The invention discovers that the expression level of the Kir4.2 protein (formed by KCNJ15 gene coding) in renal tissues of a fibrotic mouse is obviously lower than that of a wild type mouse for the first time, and the difference has statistical significance; moreover, after the Kir4.2 is specifically knocked out of the renal proximal tubule, renal fibrosis under a UUO and FA induction model can be aggravated, and the fibrosis process of human renal tubule epithelial cells induced by TGF-beta1 in vitro can be remarkably reversed by overexpression of the Kir4.2. Therefore, Kir4.2 can be used as a drug, a drug target or a target gene in gene therapy, is applied to prevention, alleviation and treatment of renal fibrosis, and can provide a new strategy for prevention and treatment of renal fibrosis. The invention further provides a construction method of the kidney proximal tubule specificity Kir4.2 knockout animal model, and the constructed animal model can be used for researching renal fibrosis pathogenesis and screening drugs for preventing, relieving or / and treating renal fibrosis and has a wide application prospect.

The application relates to the technical field of animal model construction, and particularly discloses a Sypl1 gene knockout colorectal cancer mouse model construction method and application, which comprises the following steps: synthesizing a specific target site gRNA of a Sypl1 gene, mixing Cas9 protein and the target site gRNA to obtain an injection compound, microinjecting the injection compound into mouse zygotes, and transplanting the surviving zygotes into the oviducts of pseudopregnant female mice to obtain F0 generation mice, and the mice born after 20 days are Sypl1 gene knockout mouse animal models; and a chemical induction modeling method is used to construct a colorectal cancer model in wild type mice and gene knockout mice. The Sypl1 gene knockout mouse is constructed by using a CRISPR / Cas mediated genome engineering technology Sypl1 combined with the most widely used colorectal cancer chemical inducer azoxymethane / dextran sodium sulfate, a new colorectal cancer mouse model is constructed, compared with a traditional chemical induction model, the tumor formation rate can be significantly improved, and the tumor formation time can be shortened, and a more efficient animal model is provided for colorectal cancer research.

The application discloses a construction method and application of a Clock gene site-directed mutant mouse model, and belongs to the technical field of basic medical animal models. The construction method comprises the following steps: gRNA vector design and construction, synthesis of a Donor Oligo sequence, co-injection of the gRNA vector, a Cas9 vector and the synthesized Donor Oligo into a fertilized egg to obtain F0 generation transgenic mice; and crossbreeding of the F0 generation mice with wild type mice to obtain F1 generation transgenic mice. Phenotype analysis on the F1 generation transgenic mice shows that the Clock gene site-directed mutant mouse exhibits an age progression heart failure phenotype, which is basically consistent with heart failure caused by aging in a natural physiological state, and therefore, the mouse can be used as a model of chronic heart failure disease, and is expected to be used for in-depth research on the pathogenesis based on the model, thereby providing technical support for prevention and treatment of the disease.

The invention relates to the technical field of animal model construction, and particularly discloses a construction method and application of a Sypl1 gene knockout colorectal cancer mouse model, and the construction method comprises the following steps: synthesizing a specific target site gRNA of a Sypl1 gene, uniformly mixing Cas9 protein and the target site gRNA to obtain an injection compound, microinjecting the injection compound into a mouse fertilized egg to obtain a Sypl1 gene knockout colorectal cancer mouse model. And transplanting the survival fertilized ova into the salpingtube of a pseudopregnant female mouse for 20 days to obtain a mouse which is an F0-generation mouse, obtaining a Sypl1 gene knockout mouse animal model, and constructing a colorectal cancer model in a wild type mouse and a gene knockout mouse by adopting a chemical induction modeling method. A Sypl1 gene knockout mouse is constructed through a CRISPR / Cas mediated genome engineering technology, a novel colorectal cancer mouse model is constructed in combination with a colorectal cancer chemical inducer azomethane / dextran sodium sulfate which is most widely applied at present, and compared with a traditional chemical induction model, the novel colorectal cancer mouse model has the advantages that the tumor formation rate can be remarkably increased, the tumor formation time can be shortened, and the tumor formation cost can be reduced. And a more efficient animal model is provided for colorectal cancer research.

The invention discloses application of alpha1-acid glycoprotein in preparation of a medicine for treating lung injury caused by acute pancreatitis, and belongs to the technical field of biological medicine. An acute pancreatitis lung injury model is constructed through ORM1 gene knockout mice, the mice are divided into a normal group, a model group and a treatment group for research, target organ pathological injury degree detection, immunohistochemical analysis and serum inflammatory factor expression quantity are performed after injection administration, and the curative effect difference of the gene knockout mice and wild type mice is compared, so that the acute pancreatitis lung injury model is established. And the treatment effect of the alpha1-acid glycoprotein on the acute pancreatitis lung injury is verified. The invention provides a key basis for research, development and preparation of drugs for treating acute pancreatitis lung injury, and has important clinical application value.

The invention relates to the technical field of biotechnology, in particular to a two-leaf aortic valve disease model mouse as well as a construction method and application thereof, and the construction method comprises the following steps: designing gRNA of a targeted mouse FGFR3 exon 5-7, adapting a T7 promoter, and carrying out transcription purification after off-target prediction; mixing gRNA with Cas9, adding glycerol, incubating to obtain an RNP compound, and verifying the activity; rNP is injected into fertilized eggs, and after culture, the fertilized eggs are transplanted to pseudopregnant female rats to obtain an F0 generation; the F0 and a wild type mouse are subjected to cage combination to obtain F1, F1 heterozygotes are subjected to reproduction to obtain F2, and homozygotes are identified through PCR and sequencing; and taking F2 heart HE staining ornamental deformity, and detecting the expression of FGFR3 by RT-PCR and WB. According to the model, R-N type two-leaf aortic valve and right ventricle double-outlet deformity can be stably reproduced, the construction method is accurate and efficient, the model can be used for disease mechanism research and drug screening, and the blank of specific subtype research tools is filled.