42 results about "Pig model" patented technology

The invention discloses a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 system and application thereof to construction of swine-derived recombinant cells with insulin receptor substrate gene defects. The invention provides an sgRNA (ribonucleic acid) combination which is composed of sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. The invention provides a plasmid combination. The plasmid combination is composed of four plasmids, and the four plasmids are respectively subjected to transcription to obtain sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. Theapplication of the sgRNA combination or the plasmid combination is as follows: preparation of recombinant cells; preparation of a diabetic cell model; preparation of a diabetic animal model. Accordingto the invention, sgRNAIRS1-1 is as shown in SEQ ID NO:8; sgRNAIRS1-3 is as shown in SEQ ID NO:10; sgRNAIRS2-2 is shown as SEQ ID NO:14; and sgRNAIRS2-3 is as shown in SEQ ID NO:15. A solid foundation is laid for the preparation of diabetic pig models, and important application value for the research and development of diabetic medicaments is achieved.

The invention provides a heavy immunodeficiency pig model and a construction method and application thereof. The method utilizes a BE3 point mutation tool for knocking out RAG1, RAG2 and IL2RG genes to obtain an immunodeficiency models; due to the C mutation T point mutation tool, the multi-gene knockout immunodeficiency model is obtained, the safety is better, the immunodeficiency degree is higher, T cells, B cells and NK cells are completely lost, follow-up application research is facilitated, the construction method is simple and efficiency, and the wide application prospect and huge marketvalue are achieved.

The present invention provides transgenic, large non-human animal models of diseases and conditions, as well as methods of using such animal models in the identification and characterization of therapies for the diseases and conditions.

The present invention provides transgenic, large non-human animal models of diseases and conditions, as well as methods of making and using such animal models in the identification and characterization of therapies for the diseases and conditions.

The invention belongs to the technical field of biology, and specifically discloses a targeting vector for pig GHR gene knock-out and applications thereof. The targeting vector is a sgRNA expression vector based on a CRISPR / Cas system; wherein the target sequences of sgRNA are arranged on the 10th exon and 3'UTR of a pig GHR gene. The provided targeting vector is applied to targeting knock-out ofpig GHR gene, and the knock-out efficiency can reach 12.7%. After the GHR gene is successfully knocked out, somatic cells can be used to establish a GHR gene knock-out pig model by a somatic cell nuclear transplanting technology, and materials are provided for the research of physiological action and molecular mechanism of GHR gene.

The invention provides a method for establishing a coronary artery microvascular spasm pig-model, and belongs to the technical field of medical models. The method comprises that after bilateral femoral arteries of a test pig are free, distal ends of the femoral arteries are ligatured by wires, a left-side sheathing canal is fed into a pigtail catheter, blood flow dynamics indexes are determined, in a right-side sheathing canal, a coronary artery radiography process is carried out so that the middle of an anterior descending branch is displayed, and a neuropeptide Y is injected into the coronary artery so that the coronary artery microvascular spasm pig-model is obtained. The coronary artery microvascular spasm pig-model has good singularity, well fits to the clinical disease fact, satisfies clinical psychological stress characteristics, can be operated simply, is scientific and reasonable, has short modeling time, small damage and low animal mortality, realizes coronary artery microvascular evaluation on the carrier, realizes qualitative and quantitative evaluation of microvascular spasm state and degree, realizes real-time observation, provides a novel research means for basic clinical research and is suitable for popularization application.

The invention belongs to the field of animal medical models and particularly relates to a method for building a small pig model for researches on lower limb deep venous thrombus formation and pulmonary embolism. The method is characterized in that the method comprises the steps of firstly forming small pig posterior limb deep venous thrombus, cutting a small pig anterior limb cephalic vein open for intubation, injecting the in-vitro prepared thrombus into a superior vena cava and delivering the thrombus to a pulmonary artery through cardiac circulation to cause pulmonary artery embolism to build the small pig model for researches on lower limb deep venous thrombus formation and pulmonary embolism, wherein aminomethylbenzoic acid injection in appropriate proportion being 1: (15-20) is added during the preparation of the in-vitro thrombus. By building the small pig model for researches on lower limb deep venous thrombus formation and pulmonary embolism, a good animal model can be provided for pathophysiologic researches on diseases, clinical drug researches and surgical practices. The method provided by the invention has the advantages of simple anesthesia operation, clear effect, simple and rapid surgical operation, high pulmonary embolism formation success rate, low fatality rate, convenient inspection means and the like.

The present invention relates to a genetically modified pig as a model for studying psoriasis. The modified pig model displays one or more phenotypes associated with psoriasis. Disclosed is also a modified pig comprising a mutation in the endogenous ILK-I Ra, JunB / cJun, CD18, IKK2, and / or LIG1 gene, and / or a human, porcine and / or murine PPARs, PPAR-δ, lκB-α, STAT3c, Integrin beta 1, Integrin alpha 2, MEK1, Amphiregulin, BMP-6, VEGF, JunBΔec-JunΔep, IL-I a, TGF.beta 1, CD18 hypo, Cre / lkk2FL / FL, Dsg1, SCCE, TGF-a, TNF-a, IL-20, IFN-g, LIG1 KO, KGF, IL-6, PAFR1 Cre / lkk2FL / FL, IL1 R, Dsg3, IFN-gamma, p40, ILI Ra, IKK2, JunB / c-Jun, and / or LIG1 gene, transcriptional and / or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of psoriasis, for screening the efficacy of a pharmaceutical composition, and a method for treatment of human being suffering from psoriasis are disclosed.

The present invention relates to a genetically modified pig as a model for studying breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex. The modified pig model displays one or more phenotypes associated with any of said disorders. Disclosed is also a modified pig comprising a modified endogeneous BRCA1 and / or BRCA 2 gene, and / or a modified ornithine transcarbamylase gene, and / or a modified Keratin 14 gene and / or a transcriptional or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex; methods for screening the efficacy of a pharmaceutical composition; and a method for treatment of a human being suffering from breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex are disclosed.

The invention belongs to the technical field of medical preparations of inorganic compounds containing ammonia, in particular to a construction method of a feces transplantation experiment piglet model, and the method is obtained by performing feces bacteria transplantation on a sterile newborn piglet. The method of the invention avoids the treatment of antibiotics or other sterilization drugs, the constructed fecal bacteria transplantation pig model has good effect with no drug residue, and the function of donor flora can be reflected more truly and reliably; the expression level of tight junction protein ZO-1 and Occludin in the empty intestine of the piglet can be improved; the development of muscular layer and gland of the intestinal wall of the piglet can be promoted; and the contentof various volatile short-chain fatty acids such as acetic acid, n-propionic acid, isobutyric acid, n-butyric acid, isovaleric acid, n-valeric acid and the like in intestinal tracts of the piglet is improved.

The invention discloses an in-vitro construction method of a pig liver tissue organoid model. The method comprises the steps of shearing and cleaning the liver tissues of a newborn piglet, and addingdigestive juice for incubation and digestion to obtain organoid fine particles; mixing the organoid fine particles with matrix gel, and performing inoculation; and after the matrix gel is solidified,performing incubation for 4 days by using an induction culture medium, then performing culture by using an expansion culture medium, and replacing the expansion culture medium once every 3-4 days, soas to obtain the pig liver tissue organoid model. The construction method is simple, rapid in construction and high in operability; the organoid model is constructed directly through the organoid fineparticles of a newborn animal; the organoid model not only can truly reflect the internal environment in a living pig model, but also can also be used for studying liver growth and lipid metabolism in the living pig model at the molecular cell level; and sampling of living pigs is reduced.

The invention discloses an intelligent oestrus testing boar machine comprising a platform. Two symmetrical driven wheels are arranged at the front part of the platform; a driving mechanism for drivingthe platform to move and turn is arranged at the tail part of the platform; the driving mechanism includes a worm gear steered by a controller; the worm gear is hinged to the platform; a rack is fixedly connected in the worm gear; a driving wheel is mounted on the rack and is driven by a first motor; and a worm is driven by a second motor. The platform is provided with a swing plate through a spring and connected with an eccentric wheel through a connecting rod; and the eccentric wheel is connected with the second motor through a clutch mechanism. A container for storing boar odor liquid is arranged at the opening of a boar model. The first motor and the second motor are both positive and negative rotation motors. The boar model on the platform is transported to the front of a to-be-checked sow through the driving mechanism; and the volatilization of boar odor liquid is used for testing whether the sow is in an oestrus or not. Therefore, the problems of high boar feeding cost and highoestrus checking cost are solved.

The present invention relates to a genetically modified pig as a model for studying breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex. The modified pig model displays one or more phenotypes associated with any of said disorders. Disclosed is also a modified pig comprising a modified endogeneous BRCA1 and / or BRCA2 gene, and / or a modified ornithine transcarbamylase gene, and / or a modified Keratin 14 gene and / or a transcriptional or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex; methods for screening the efficacy of a pharmaceutical composition; and a method for treatment of a human being suffering from breast cancer, mitochondria related protein folding disorders and / or epidermolysis bullosa simplex are disclosed.

The invention discloses shRNA (short hairpin ribonucleic acid) capable of inhibiting IRS (insulin receptor substrate) 2 gene expression and application thereof, belonging to the technical field of gene engineering. The nucleotide sequence of the shRNA is shown in SEQ ID No.1. The invention also provides an interference vector of shRNA capable of inhibiting IRS 2 gene expression. Through cloning the compared and analyzed pig source IRS 2 gene conserved sequence, interference fragments with restriction enzyme cutting site BamH I and Hind III are synthesized, the interference fragments are connected onto a plasmid vector linearized by BamH I and Hind III, and effective interference vectors are screened out. The shRNA and the interference vector thereof can effectively inhibit IRS2 gene expression, affects cell glucolipid metabolism, and lays a foundation for construction of type 2 diabetic pig model.

The present invention provides a production method and applications of an autosomal dominant polycystic kidney disease gene mutation pig, wherein a gene editing technology is used to introduce insertion mutation into the 5# exon of the pig PKD1 gene and knockout the pig PKD1 gene to construct an autosomal dominant polycystic kidney disease gene mutation pig model. The constructed gene mutation pig model of the present invention can be used for research and drug screening of the autosomal dominant polycystic kidney disease.

The invention relates to the field of medicine, in particular to a pulmonary arterial hypertension non-human primate model and a construction method thereof.The construction method of the pulmonary arterial hypertension non-human primate model comprises the following steps that a Seldinger technology is used for puncturing femoral veins to establish a right heart catheter channel; the degradable embolism particles or the degradable embolism particle preparation is injected into the right atrium of an experimental animal body through a right heart catheter, the degradable embolism particles block pulmonary artery branches and induce autologous thrombus formation through blood circulation, and therefore the pulmonary arterial hypertension animal model is obtained. Compared with a rodent model or a pig model, the pulmonary arterial hypertension non-human primate model is closer to a disease state under human pulmonary arterial hypertension; the degradable embolism particles used by the model can be degraded and absorbed along with time in the animal body, and the thrombolytic drug can obviously accelerate the process, so that the model can be used for screening the thrombolytic drug, and is suitable for developing thrombolytic drug curative effect evaluation experiment research.

The invention discloses a construction method and application of an hACE2 humanized transgenic pig. A CRISPR-Cas9 gene editing technology is adopted, hACE2cDNA is used for replacing an ACE2 gene of a pig, and the problem that a pig model cannot well reproduce human disease characteristics due to the fact that pig endogenous ACE2 is also expressed while humanized ACE2 is expressed is solved. The hACE2 humanized pig model constructed by the invention is an ideal large animal pig model capable of simulating clinical symptoms and treatment response of human COVID-19, can be used for research on SARS-CoV-2 infection and pathogenesis, and is particularly suitable for research on multi-organ diseases, severe diseases, chronic diseases and sequelae caused by SARS-CoV-2 infection, the model can be used for development and testing of diagnostic technology, antiviral therapy and vaccines and evaluation of curative effect and safety of candidate drugs, and is beneficial to solving the problem of lack of an effective new coronal pneumonia animal model at present.

The invention discloses preparation and application of a coral snake bitten pig model. Coral snake venom freeze-dried powder is injected into pig muscles, the coral snake bitten pig model is established, and the defect that no experimental animal model exists for coral snake bites is overcome. The model is close to the characteristic that the disease course of human beings bitten by coral snakes is long, can simulate the coral snake bite process, can be used for researching clinical characterization, pathophysiological mechanism and precise treatment of the incidence of coral snake bites, andcan finally evaluate the curative effects of anti-snake venom serum and anti-venomous snake bite drugs.

The invention discloses a manufacturing method of a Chinese cobra bite pig model and half lethal dose (LD50) of cobra venom to pigs and application of the cobra venom to an animal model. Cobra venom freeze-dried powder is dissolved and then injected into pig leg muscles according to 2mg / kg, and a target animal model can be obtained after 6 hours, so that the problem that no Chinese cobra bite model of large animals in the prior art is solved. The model has the advantages of high success rate, strong repeatability, stable pathological change and the like. The cobra bite disease condition is long, the model simulates human being bite by the cobra, the physiological and pathological dynamic process of the human being bite by the cobra can be well close to, and dynamic analysis of the disease clinical manifestation and mechanism of the cobra bite is facilitated. The model can provide a good animal model for researching metabolic change of Chinese cobra venom in a human body and a muscle necrosis mechanism, and provides an innovative technical method support for researching clinical characterization of cobra bite, pathophysiological mechanism, precise treatment, screening of anti-venomous snake bite medicines, evaluation of curative effect of anti-venom serum and the like.

The invention discloses a CRISPR / Cas9 system for constructing Leber congenital amaurosis cloned porcine nuclear donor cells and application of the CRISPR / Cas9 system. The sequence of the gRNA of the pig RPE65 gene is shown as SEQ ID NO. 23. The CRISPR / Cas9 system for editing the pig RPE65 gene comprises the gRNA expression vector disclosed by the invention and a Cas9 expression vector. Wherein theCas9 expression vector is pU6gRNA eEF1a-mNLS-hSpCas9-EGFP-PURO. Gene editing is carried out by adopting the Cas9 efficient expression vector which is combined modified by gRNA screened by the invention, and the editing efficiency is improved by over 100 percent compared with that of an original vector. The invention lays a solid foundation for the preparation of a Leber congenital amaurosis pig model, and has important application value for the treatment and pathological research of the Leber congenital amaurosis pig model.

The invention provides a method for constructing a low-backfat-thickness pig model. The method comprises the following steps: S1, selecting healthy piglets at ages of 4-5 months; S2, adopting basal diet to adaptively feed piglets for 3-7 days; S3, continuously feeding the piglets for 20-30 days by high-calcium diet to obtain the low-backfat-thickness pig model, wherein the mass percentage of calcium in the high-calcium diet is 0.7%-4.0%. The composition and proportion of the basal diet are adjusted, for the high-calcium diet, mineral meal is added on the basis of the composition of the basal diet to control the content of Ca in the high-calcium diet, so that accumulation of subcutaneous fat is reduced, backfat thickness is reduced, the lean meat percentage is increased, and the method hasthe advantages of being low in cost and convenient to operate. The method can be applied to the pig breeding industry, and lays a technical foundation for production of tender, tasty and fine pork products with a high lean meat rate.

Human and humanized monoclonal antibodies which binds specifically to subunit A of Shiga like toxin II have been developed which are effective to prevent or ameliorate one or more symptoms of HUS in a human. Effective dosages for treatment or prevention range from approximately 0.1 to 5.0 mg of antibody / kg of patient weight. The examples demonstrate the preferred dosage ranges based on the pig model, and what is being tested in phase I clinical trials. Antibodies are preferably transfused over a period of two hours, although this will depend on the patient and the disease state at the time of treatment. Preferred dosages for treatment of humans are between 0.1 mg / kg-5.0 mg / kg of 5C120, or an equivalent dosage of another antibody to subunit A of STX2. In the most preferred embodiments, dosages of 0.1 mg / kg, 0.5 mg / kg, or 5.0 mg / kg of 5C12 (low dose, anticipated therapeutic dose based on animal data and high dose) are administered.

The present invention relates to a transgenic pig as a model for studying a cone affecting disease, in particular a cone dystrophy or cone-rod-dystrophy, wherein the pig model expresses a dominant negative guanylate-cyclase-2D (GUCY2D) protein, in particular a GUCY2D protein comprising at least one mutation responsible for the appearance of a CORD6 cone dystrophy in a human being. The invention further relates to methods by which the transgenic pig is produced, to uses of said transgenic pig or of one of its elements to identify new biomarkers of a cone affecting disease and / or new compounds for preventing or treating such a disease. Novel methods for preventing or treating a cone affecting disease or for evaluating conditions needed to alleviate such a disease are further herein described.

The invention relates to a method for constructing a severe immunodeficiency and liver injury dual pig model and application, and belongs to the technical field of animal biology. According to the method, RAG2, IL2RG and FAH genes in porcine fetal fibroblasts are knocked out by adopting a CRISPR / Cas9 technology, RAG2- / - / IL2RG- / Y / FAH- / -three-gene editing cloned pigs are constructed by utilizing a somatic cell nuclear transplantation technology, and the severe immunodeficiency and liver injury dual pig model is obtained through phenotype analysis and identification. The problems of long production cycle, low efficiency, irreversible injury, lack of immunodeficiency and non-ideal humanization degree of simultaneous application of liver injury and the like in the existing model construction technology are overcome, and batch construction of the severe immunodeficiency and liver injury dual pig model can be realized through a continuous cloning technology, and the method has huge advantages and potential market application prospects in the related fields of tumor biology, cell transplantation, humanized animal models and the like.

The invention relates to a pig weight estimation method, system and device and a storage medium. The method comprises the steps of constructing a pig model through three-dimensional simulation, and then constructing a large number of two-dimensional projection pictures and three-dimensional pig weight data in a projection mode so as to train a deep learning model; acquiring a two-dimensional picture of the pig acquired by the camera; and inputting a pre-trained neural network model according to the parameters of the camera, the variety of the pig and the two-dimensional picture to obtain an estimated value of the weight of the pig. According to the pig weight estimation method provided by the embodiment of the invention, the estimated value of the pig weight can be accurately obtained through the two-dimensional picture of the pig by adopting a machine learning method under the condition of different pig varieties and camera parameters, and the efficiency of pig breeding is improved.