8 results about "Mouse strain" patented technology

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 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.

This invention discloses a gRNA combination and its application in the preparation of drugs for the prevention of metabolic-associated fatty liver disease (MASLD). The gRNA combination, when mixed with Cas9 protein, yields an RNP complex, which, when microinjected into mouse zygotes, can breed a stable and heritable mouse strain with TMEM68 gene knockout. Combining lipidomics, transcriptomics, and primary hepatocyte functional verification, the core regulatory role of TMEM68 in hepatic lipid metabolism is revealed for the first time systematically. Experiments demonstrate that TMEM68 deficiency significantly reduces the storage of triglycerides (TAG) in the liver and hepatocytes, decreases lipid droplet formation, and reshapes the metabolic homeostasis of various lipids such as glycerophospholipids, cholesterol esters, and bile acids. Therefore, the gRNA combination of this invention, or the RNP complex obtained by mixing the gRNA combination with Cas9 protein, can be used to prepare drugs for the prevention of MASLD, providing a novel intervention strategy for MASLD prevention with broad application prospects.

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 present invention discloses the application of siRNA targeting the mGluR2 gene, monoclonal antibodies against the extracellular domain of the protein encoded by the mGluR2 gene, truncated peptides containing the extracellular domain of the protein encoded by the mGluR2 gene, or recombinant vectors containing the gRNA gene sequence targeting the mGluR2 gene and the CAS9 protein gene sequence in the preparation of preparations for inhibiting influenza virus, swine acute diarrhea syndrome coronavirus, porcine epidemic diarrhea virus, Japanese encephalitis virus, and porcine reproductive and respiratory syndrome virus. The present invention also discloses the application of a genetic engineering operation method for knocking out the mGluR2 gene in the preparation of a new mouse strain with enhanced influenza resistance. The aforementioned applications have prospects for promotion.

The invention discloses a construction method and application of an animal model with insufficient bladder activity and fibrosis, and belongs to the technical field of biology. The construction method comprises the following steps: constructing a Krt20-Cre-tdTomato mouse strain Krt20-Cre-tdTomato through a gene editing technology, and specifically, inserting a Cre-2A-tdTomato-Wpre-pA sequence into a first exon of a Krt20 gene by adopting a CRISPR / Cas9 mediated homologous recombination technology, and constructing the Krt20-Cre-tdTomato mouse strain which can be used as an animal model with insufficient bladder activity and fibrosis. The method is applied to the research of screening medicines for treating bladder activity deficiency and bladder fibrosis. According to the method, the problems that the existing lower urinary tract symptom modeling process is complicated and the model types are insufficient are solved, and the constructed bladder activity deficiency and fibrosis animal model can provide a basis and a preclinical animal model for screening potential drug targets.

The present invention discloses the use of a single-nucleotide polymorphism (SNP) marker for identifying inbred mouse strains and primer sequences. The SNP marker provided by the present invention includes multiple or all of eight SNP sites and can identify some or all of ten inbred mouse strains selected from DBA / 2, FVB, BALB / cJ, C57BL / 6, C57BL / 6-bg, NPI, NCPC / 2, NU / J, C3H, and T739, thereby improving the scope and efficiency of inbred mouse strain identification.

Inventors have used FXYD7 knock-out mouse line and DRG-specific conditional Knock-out mice to demonstrate that the constitutive or DRG-specific knock-out of the FXYD7 gene specifically alleviate mechanical chronic pain induced by peripheral inflammation, but not by peripheral nerve lesions. They have also showed that: i) FXYD2 and FXYD7 are expressed by distinct and complementary somatosensory neuron subtypes suggesting specific functions in the somatosensory system; ii) FXYD7 is largely dispensable for neuronal differentiation and survival; iii) in FXYD7− / − animals, FXYD7 expression was not affected in the DRG. Accordingly, the present invention relates to a method for treating a subject suffering from a chronic pain disorder comprising a step of administrating said subject with a therapeutically effective amount of an inhibitor of FXYD7.