262 results about "Mus spretus" patented technology

The invention discloses a construction method and application of an NMD escape model of an Alport syndrome mouse. A non-human animal model carrying Col4a5 gene c.4432delG frame shift mutation is prepared on the basis of a CRISPR / Cas9 gene editing technology. The method comprises the following steps: co-injecting gRNA of a 49 exon of a targeted Col4a5 gene, homologous recombinant donor oligonucleotide containing c.4432delG mutation and Cas9 nuclease into a mouse fertilized egg, and carrying out embryo transplantation to obtain an F0-generation mutant mouse; a mutation site is verified by combining PCR (Polymerase Chain Reaction) with sequencing, and a stably inherited mutation line is established through two generations of breeding. Through verification, the model accords with pathological characteristics of the Alport syndrome, can stably simulate typical clinical manifestation and pathological characteristics of the human X-linked Alport syndrome, and can be used as an important tool for research of the Alport syndrome.

The invention belongs to the technical field of gene engineering, and relates to a construction method of a mouse model capable of rapidly degrading MSTN protein, which comprises the following steps: sequentially inoculating a 5'homologous arm, 3xEAAAK, an FKBP1A gene synonymous mutation sequence, 3xEAAAK, a red fluorescent protein mCherry sequence and a 3 'homologous arm on a Donor vector to obtain a Donor recombinant vector; designing gRNA based on a third exon of the Mstn gene, mixing the gRNA, Cas9 protein and a Donor recombinant vector, injecting the mixture into a mouse fertilized egg, and performing in-vitro culture and transplantation to obtain an F0-generation mouse; and hybridizing the F0-generation mouse with a wild type, or continuously breeding the filial generation to obtain the knock-in mouse model. According to the invention, a dTAG targeted protein degradation technology is utilized, the obtained homozygous knock-in mouse is subjected to dTAG administration, and rapid and reversible degradation and removal of MSTN protein can be realized.

The invention relates to the technical field of animal experiment equipment, and discloses an isolated culture device for mouse sweet water preference test in an IVC cage, which comprises a shell cover, the partition plate is fixedly installed on the inner side wall of the shell cover, and an inner cavity of the shell cover is divided into a living area and a functional area through the partition plate; the sliding cover is arranged in the living area of the shell cover and used for containing solid feed, and multiple sets of feeding trough openings which are arranged at equal intervals are formed in the bottom of the sliding cover; according to the device, an innovative cage-in-cage design is adopted, a single cage is converted into a plurality of independent experiment units on the premise that an existing IVC system is not changed, and cage position resources are greatly saved. The functions of isolation, double-bottle water drinking and independent feeding are integrated, and convenient operation is achieved through the pull-out type fixing plate and the sliding cover plate. The centrifugal tube is adopted to replace a special water bottle, so that the cost is reduced, the SPF environment is kept in the whole process, data accuracy and reliability are ensured, and the traditional SPT test problem is effectively solved.

The spliced sleep deprivation mouse cage comprises a plurality of square cage bodies, a tray and a food box, the square cage bodies are transparent structural members, the top of each square cage body is provided with an organism inlet and at least one square notch, the square notches are formed in the side edges of the square cage bodies, and the food box is arranged on the tray. A plurality of water leakage holes are evenly formed in the bottoms of the square cage bodies at intervals, clamping grooves are formed in the tray, the square cage bodies can be spliced and embedded into the clamping grooves, circulation gaps are formed between the bottom walls of the square cage bodies and the inner bottom wall of the tray, and a water injection opening is formed in the side wall of the tray. When the sides, provided with the square notches, of any four square cage bodies are spliced, a containing groove communicated with the four square cage bodies is formed, and the food box is arranged in the containing groove. The splicing type sleep deprivation mouse cage can effectively deprive the sleep of mice, the problems of poor sleep deprivation and wire entanglement caused by mixed feeding of the mice are solved, the water level height in each mouse cage is consistent, and meanwhile, the water quality is convenient to replace.

The invention discloses a construction method and application of a GSDMB intestinal epithelium specific knock-in mouse model. The construction method comprises the following steps: (1) designing gRNA according to an ROSA26 gene, constructing a Donor vector containing a GSDMB gene, introducing the gRNA and the Donor vector and Cas mRNA into a fertilized egg of a receptor mouse, transplanting the fertilized egg and Cas mRNA into a pseudopregnant mouse body for reproduction, and screening out a GSDMB gene knock-in F0-generation mouse; (2) mating and breeding the F0 generation mouse and a wild type mouse, and screening out an F1 heterozygote mouse with GSDMB knocked in; (3) carrying out inbreeding propagation on the F1 heterozygote mouse, and screening out an F2 homozygote mouse in which GSDMB is knocked; (4) the F2 homozygote mouse and the intestinal epithelium specific Vilin-Cre transgenic mouse are subjected to mating breeding, and a heterozygote mouse with GSDMB intestinal epithelium specific knock-in is screened out; and (5) carrying out inbreeding propagation on the heterozygote mouse with the GSDMB intestinal epithelium specific knock-in, and screening out the homozygote mouse with the GSDMB intestinal epithelium specific knock-in.

Studies of non-human species are often pursued to validate or extend the biological insights of studies exploring gene-phenotype relationships in humans. When the results of non-human studies do not align with those of human studies, various questions arise regarding the study design, contextual factors, reagents and assays employed, and other related aspects. It is arguable that outside of the experimental conditions, there could be biological reasons why non-human and human studies produce discordant results exploring gene-phenotype relationships. These reasons might include the possibility that the gene in question has evolved a different function in one of the species or that the regulatory mechanisms governing its expression have undergone changes. This study assessed the potential of certain gene features, such as sequence conservation levels and aspects of regulatory architecture, to explain the concordant and discordant results obtained from studies involving humans and mice.

The invention relates to a mouse model construction method for psoriasis research based on dihydroxyisobutyrylation modification of SBSN 600 site lysine. The mouse model construction method comprises the following steps: 1, constructing a plasmid comprising an SBSN gene sequence and a DNA fragment of which the 600 site of the SBSN sequence is mutated from lysine (K) to threonine (T); 2, culturing the mouse embryonic stem cells; step 3, carrying out electric transformation on the embryonic stem cells; 4, screening the medicines; 5, calling and culturing the embryonic stem cells screened in the step 4; step 6, injecting the embryonic stem cells called and cultured in the step 4 into a mouse blastocyst, then culturing the mouse blastocyst, and transplanting the cultured blastocyst into the uterus of a pseudopregnant female mouse, so that the mouse generated by delivery of the female mouse is a chimeric mouse; and 7, breeding the chimeric mouse and identifying the genotype of the mouse through PCR (Polymerase Chain Reaction).

The present application discloses a gene point mutation mouse model construction method, comprising: by taking T274M mutation on Kcnq2 Exon6 as a target, designing sgRNA and ssODN; selecting a female mouse of a set week age as an ovum donor, injecting a certain dose of serum gonadotropin and human chorionic gonadotropin into the abdominal cavity of the female mouse, then mating the female mouse with a mating male mouse, and collecting zygotes of the mated female mouse; injecting Cas9 protein, ssODN and sgRNA into the pronuclei of the zygotes by means of a microinjection technique, transplanting the zygotes obtained after injection into the ampulla of the fallopian tube of a surrogate female mouse, and obtaining a neonatal mouse after the pregnant female mouse gives birth; respectively designing a pair of identification primers F1, R1 at two ends of a mutation site on the basis of a Kcnq2 gene point mutation site, identifying the neonatal mouse, and screening out a Kcnq2T274M / + point mutant mouse in which T274M mutation is successfully knocked.

The invention relates to a gene point mutation mouse model construction method. The method comprises the following steps: designing sgRNA and ssODN by taking T274M mutation on Kcnq2Exon6 as a target spot; the method comprises the following steps: selecting a female mouse at a set week age as an ovum donor, injecting a certain dose of serum gonadotropin and human chorionic gonadotropin into the abdominal cavity of the female mouse, mating with a breeding male mouse, and collecting fertilized ova of the mated female mouse; injecting Cas9 protein, ssODN and sgRNA into the pronucleus of the fertilized ovum through a micro-injection technology, transplanting the fertilized ovum after injection into the oviduct pot belly of the female rat to be pregnant, and obtaining a newborn rat after the female rat to be pregnant is delivered; according to a Kcnq2 gene point mutation site, a pair of identification primers F1 and R1 are designed at the two ends of the mutation site respectively, and a newborn mouse is identified, so that a Kcnq2T274M / + point mutation mouse with T274M mutation successfully knocked in is screened out. The Kcnq2T274M / + point mutation mouse editing method is simple in preparation and high in repeatability, and a larger number of Kcnq2T274M / + point mutation mice can be obtained while the editing efficiency is improved.

The invention provides a method for establishing a mouse postoperative delirium model under a simulated high altitude urgent entry environment, and relates to the technical field of animal model establishment. According to the method for establishing the model for simulating postoperative delirium of the mouse in the sudden entering plateau environment, a control group (group C), a plateau group (group H), a POD model group (group P) and a plateau POD model group (group HP) are included, behavior changes of the mouse are evaluated through a Y maze experiment, then the mouse is killed, brain tissue is taken, and the mouse is subjected to postoperation delirium simulation in the sudden entering plateau environment. The mouse hippocampus tissue structure damage change is observed through morphological methods such as Nissl staining and a transmission electron microscope, meanwhile, blood is taken through mouse eye sockets, the concentration of inflammatory related indexes IL-6 and TNF-alpha in serum is measured through an ELISA method, and the stability and repeatability of model establishment are known. The plateau postoperative delirium mouse model prepared by the invention can generate obvious damage to brain tissues and neurological functions, and is high in operability and good in repeatability.

The invention provides a construction method and application of a WARS2 point mutation mouse model. A part of gene segments of a sixth exon of an endogenous WARS2 gene in a homologous chromosome of a mouse are humanized, and WARS2 c.799Ggt, WARS2 c.799Ggt, WARS2 c.799Ggt, WARS2 c.799Ggt, WARS2. And the C (p.Ala267Pro) site is subjected to point mutation. The construction method disclosed by the invention provides convenience for obtaining a mouse model which is similar to human WARS2 gene p.Ala267Pro pathogenic mutation, and provides a basis for researching related factors of human WARS2 A267P / A267P mutation and diagnosing and treating the human WARS2 gene p.Ala267Pro pathogenic mutation. Therefore, researchers can deeply analyze the biological function of the WARS2 gene, an effective research way and method are provided for exploring the internal relation between the WARS2 gene function and the PD pathogenesis, and a foundation is laid for related application.

A primer pair, method, and use for rapidly identifying newborn nude mice. On the basis that a deletion mutation occurs in the base G at position 337 of a coding region of the Foxn1 gene in nude mice, a primer pair introducing an enzyme digestion site is designed according to sequence characteristics. Using the genome of a mouse to be tested as a template, the Foxn1 gene is amplified according to a PCR-RFLP method. The PCR product is then digested with a SmaI restriction endonuclease, and the digested product is genotyped by agarose gel electrophoresis. In addition, a reaction system that can be combined with lateral flow technology is screened, thereby achieving instant detection of the newborn nude mice. The method has the characteristics of simple operation, short time consumption, low cost, instant detection, etc.

The invention relates to the technical field of medicines, in particular to novel application of galactooligosaccharide. The novel use of galactooligosaccharide inhibits the formation of gallstone. Main reagents and materials are galactooligosaccharide; a mouse calculus model feed; the experimental animals are derived from animal varieties and strains: mice, C57BL / 6J; sex: male; week age: 4 week age; the body weight is 16-20 g; the experimental environment is SPF environment feeding, and the feeding condition is that the room temperature is 20-26 DEG C; the relative humidity is 40-70%; automatic illumination is achieved, and light and shade are alternated for 12 h; the drinking water is RO membrane reverse osmosis sterile water; the feeding density is defaulted to be 4 per cage, and the padding is replaced once a week; the experimental process comprises the following steps: starting an experiment after experimental feeding of all mice for one week, and randomly dividing 36 5-week male C57BL / 6J mice into three groups, namely 12 mice in each group. The invention provides a new application of galactooligosaccharide, which can inhibit the formation of gallstone and provides a new thought for preventing the occurrence of the gallstone.

The application relates to the technical field of animal activity behavior monitoring, and discloses an animal behavior activity monitoring device, which comprises a supporting seat, supporting columns are symmetrically arranged at the bottom end of the supporting seat, a supporting frame is fixedly connected to the top end of the supporting seat, a monitoring body is arranged at the top end of the supporting frame, a receiving cavity is arranged in the supporting seat, a discharging pipe, which is in communication with the receiving cavity, is arranged at the bottom end of the supporting seat, an annular track is arranged at the top end of the supporting seat, an opening is arranged at the bottom end of the annular track, and a supporting and discharging assembly is arranged in the opening; the two feeding frames can be driven to move in the annular track, the feeding frames can push the mice to move to achieve sleep deprivation of the mice, the problem that, when a traditional feeding frame with a cuboid structure is fixedly arranged, the mice are driven to move only by a deprivation rod and the sleep deprivation of the mice is interrupted due to a gap caused by the deprivation rod is solved, and the sleep deprivation of the mice can be realized continuously.

The invention belongs to the technical field of animal model construction, and particularly relates to a mouse model based on alpha-synuclein mutation and a construction method and application thereof. The mouse Snca gene is subjected to site-specific modification and mutates at the site S129A, S129D or Y125F, an alpha-syn mutation animal model which does not need exogenous induction and can naturally develop typical PD pathology and behavior phenotypes is constructed, and the limitation of an existing model in the aspects of pathology reduction degree and experiment controllability is broken through.

The application discloses a mouse and Mongolian gerbil universal immunization sampling fixing device, and relates to the field of experimental animal fixing devices.The device comprises a supporting base, a bearing support is arranged on the front end surface of the supporting base, a sampling fixing plate is rotatably arranged on the outer surface of the bearing support, guide sliding grooves are symmetrically arranged on the upper surface of the sampling fixing plate, movable blocks are movably arranged in the guide sliding grooves, and a limiting sleeve is fixedly arranged on the front surface of the sampling fixing plate and extends from one end of the movable blocks.The synchronous transmission structure of the supporting disc, the first connecting plate and the second connecting plate drives the movable blocks on the two sides to synchronously move towards each other or in the opposite direction along the guide sliding grooves, and the movable blocks are matched with the positioning support plates which can slide in the limiting sleeve, so that the horizontal spacing and the longitudinal extension length of the limb fixing points can be simultaneously adjusted, the adjustment range completely covers the size difference of adult mice and adult Mongolian gerbils, and two special fixing devices are not needed, so that the universality of the device is improved compared with the prior art.

The invention discloses a construction method of a COL4A5-K229X point mutation X-linked Alport syndrome mouse model. The construction method comprises the following steps: aiming at c.685Agt of a No.12 exon of a mouse COL4A5 gene; carrying out T point mutation, and designing and preparing Cas9 mRNA, gRNA and a donor vector; the components are mixed and then microinjected into fertilized eggs of a C57BL / 6J mouse to obtain an F0-generation mouse; identifying the genotype through PCR (Polymerase Chain Reaction) amplification and Sanger sequencing, and screening positive mice; mating the positive F0-generation mice with the wild-type mice, and breeding F1-generation and subsequent generations; the phenotype of the model is further verified through qPCR, biochemical analysis, light microscopic examination, transmission electron microscope and immunofluorescence. The model constructed by the invention shows hematuria, proteinuria, azemia, podocyte loss and irregular thickening and layering of glomerular basement membrane, is consistent with phenotypes of human XLAS patients, and provides an animal model tool for analyzing pathogenesis and developing treatment strategies.

The invention discloses a construction method and application of an enterovirus 71 type susceptible transgenic mouse model, and belongs to the technical field of biology. The method comprises the following steps: up-regulating or enhancing or improving the content or / and activity of placenta specific protein 8 in a receptor mouse to obtain a mouse overexpressing the placenta specific protein 8, hybridizing the mouse overexpressing the placenta specific protein 8 with a wild mouse to obtain an F1-generation heterozygote mouse, a homozygote mouse obtained after the F1-generation heterozygote mice are hybridized with each other is an enterovirus 71 type susceptible transgenic mouse model. The application has the advantages that not only is the susceptibility of mice to EV71 increased, but also a proper animal model is provided for exploring the important function of Plac8 in the EV71 infection process.

The application discloses a method for constructing a CRBN gene humanized mouse model, which comprises replacing a knockout region of a mouse CRBN gene with a knock-in region of a human CRBN gene, so that the CRBN gene humanized mouse model is constructed, and the CRBN gene of the CRBN gene humanized mouse model is subjected to amino acid substitution of S369C, V380E, I391V and E431D. The application replaces a key functional domain of a mouse endogenous CRBN gene with a corresponding sequence of a human CRBN gene, so that the mouse can express a functional human CRBN protein, thereby solving the problem of lack of sensitivity of a wild-type mouse to a CRBN-dependent drug. The model provides an ideal preclinical experimental platform for studying the efficacy, toxicity and mechanism of action of a CRBN-related drug.

Non-human animals, and methods and compositions for making and using the same, are provided, wherein said non-human animals comprise a humanization of an endogenous cluster of differentiation (CD) gene, in particular a humanization of a CD47 gene. Said non-human animals may be described, in some embodiments, as having a genetic modification to an endogenous CD47 gene so that said non-human animals express a CD47 polypeptide that includes a human portion and a non-human portion (e.g., a murine portion).

The invention discloses a construction method of a glomerular IgA deposition mouse model, and relates to the technical field of animal model development and glomerular IgA dyeing. Comprising the following steps: obtaining Cas9 mRNA (messenger ribonucleic acid), guideRNA (ribonucleic acid) and donor DNA (deoxyribonucleic acid) in an in-vitro transcription manner; the method comprises the following steps: carrying out microinjection on Cas9 mRNA (messenger ribonucleic acid), gRNA (guide ribonucleic acid) and donor DNA (deoxyribonucleic acid) into fertilized eggs of a C57BL / 6J mouse; carrying out genotype identification on the F0 generation of birth mice through PCR (Polymerase Chain Reaction) amplification and sequencing; mating the F0-generation positive mouse with a wild type C57BL / 6J mouse, and breeding to obtain an F1-generation mouse; and spontaneous glomerular IgA deposition appears in 24 weeks after the F1 generation heterozygote mouse is born. According to the method, accurate mutation is introduced by utilizing a homologous recombination repair mechanism to construct a glomerular IgA deposition model, the efficiency is high, and the off-target risk is low.

The invention discloses a construction method and application of an animal model for HPV (human papillomavirus) pathogenic mechanism research, and belongs to the technical field of animal models. According to the construction method of the HPV fixed-point integration animal model, HPV16 gene knock-in is carried out on the Col1a1 gene locus of a C57BL / 6J mouse through CRISPR / Cas mediated genome engineering. The method comprises the following steps: (1) designing specific gRNA; (2) constructing a knock-in vector Donor vector; and (3) uniformly mixing Cas9 protein, gRNA and a targeting vector Donor vector to obtain an injection compound, performing micro-injection on the injection compound into a mouse fertilized egg, and sending the injection compound back to a pregnant mouse fallopian tube to cultivate offspring so as to generate a gene knock-in mouse. The animal model prepared by the method can be used as a contrast of a cervical cancer precancerous lesion animal model for scientific research.

The invention provides a premature ovarian insufficiency (POI) animal model and a construction method thereof, and particularly, the method utilizes chronic unpredictable pressure stress to construct different pathological stage models of mouse POI, and comprises the following steps: screening experimental animals with normal ovarian functions, adaptively feeding, applying various pressure stresses to the experimental animals, and obtaining the POI animal model. The POI model has a compensatory period, and is more in line with the clinical pathological process of patients suffering from premature ovarian insufficiency caused by common pressure in modern society.

The invention discloses construction and application of a TAGLN2 gene defect type mouse model, which utilizes a CRISPR / Cas9 gene editing technology, uses gRNA to guide Cas9 protein to respectively shear at specific sites at the upstream of a second exon and the downstream of a fifth exon of a TAGLN2 gene, and realizes TAGLN2 gene knockout. The mouse model constructed by the invention provides a basis for researching the molecular mechanism of the TAGLN2 gene, and is beneficial to researching the pathogenesis of metabolic diseases.

The invention discloses primers and a method for detecting Corynebacterium bovis and application of the primers and the method in detection of pathogenic bacteria of immunodeficient mice. The primers comprise an upstream primer and a downstream primer aiming at 16S rRNA (ribosomal Ribonucleic Acid) genes of the Corynebacterium bovis. The invention establishes a qPCR molecular detection method for the pathogenic bacteria Corynebacterium bovis of the immunodeficient mice, and the qPCR molecular detection method has the advantages of strong specificity, high sensitivity, high detection efficiency and the like, and can quickly and accurately detect the Corynebacterium bovis in the immunodeficient mice and the environment. The establishment of the detection method is beneficial to timely discovery and control of the bacterium, and has important significance for guaranteeing the environmental safety of personnel and the health of experimental animals. In future experimental animal quality control and related scientific research work, the qPCR detection method is expected to be widely applied, and powerful technical support is provided for further deep research on biological characteristics and pathogenic mechanisms of corynebacterium bovis.

1. Name of the product in this design: Mouse tail blood collection fixation device. 2. Purpose of this design: Used as a fixation device for blood collection from the tail of mice. 3. The key design feature of this product is its shape. 4. The image or photograph that best illustrates the design's key points: 3D view 1.

The application discloses a mouse vitiligo induction model based on chronic stress factors, wherein different kinds of stress factors capable of triggering stress response of animals are applied to stimulate the mouse periodically within a certain time period, so as to prevent the mouse from producing habitual response to the stress source and to promote the mouse to become white and the tail epidermis to be flaky. The technical method is different from the existing vitiligo model method, is based on the mechanism of occurrence and development of vitiligo observed in the clinic, and promotes the mouse to truly develop the vitiligo phenomenon from the whole instead of simulating the disease evolution locally. Compared with the existing vitiligo promoting mechanism, the effect is more stable and the authenticity is higher. The technical method is based on the objective fact that the clinical vitiligo patients often combine anxiety and depression to design the stress factors, and considers that the stress factors and the occurrence and development of vitiligo may be inter-caused and viciously circulate, namely, the adverse life events as the stress factors cause the damage and dysfunction of the melanin synthesis system at the micro level and form the vitiligo change at the macro level.

The invention discloses a method for detecting mouse bronchial Bordetella by using a qPCR (quantitative polymerase chain reaction) technology instead of a traditional sacrificial sentinel mouse, and a specific primer and probe combination comprises a forward primer sequence: 5 '-CGTTGTCGGCTTTCGTCTG-3', as shown in SEQ ID NO.1; the sequence of a reverse primer is 5 '-ACTATACGTCGGAAATCTGTTTG-3', and the sequence of the reverse primer is as shown in SEQ ID NO. 2; and a probe sequence: 5 '-FAM-CGGGCCGATAGTCAGGGCGTAG-BHQ1-3', and the probe sequence is as shown in SEQ ID NO. 3. The method disclosed by the invention has the beneficial effects that an analysis method suitable for detecting the bronchial Bordetella in the animal cage environment sample is established, the bronchial Bordetella in the environment sample can be rapidly and accurately detected and quantified by the method, and a technical support is provided for pathogen pollution control.

The application relates to a mouse model construction method for psoriasis research based on SBSN 600 lysine dihydroxyisobutyryl modification, which comprises the following steps: first step, constructing a plasmid comprising a DNA fragment of a SBSN gene sequence, wherein 600 sites of the SBSN sequence are mutated from lysine (K) to threonine (T); second step, culturing mouse embryonic stem cells; third step, electrically transforming the embryonic stem cells; fourth step, drug screening; fifth step, culturing the embryonic stem cells screened in the fourth step; sixth step, injecting the embryonic stem cells cultured in the fourth step into mouse blastocysts, then culturing the mouse blastocysts, and transplanting the cultured blastocysts into the uterus of a pseudopregnant female mouse, wherein the mouse delivered by the female mouse is a chimera mouse; and seventh step, breeding the chimera mouse and identifying the mouse genotype through PCR.

The invention relates to the field of constant-temperature heating, and particularly discloses an automatic constant-temperature heating system for tails of rats and mice, which comprises a temperature sensing module for collecting tail temperature data in real time, an environment compensation module for generating heating compensation parameters based on a Fourier transform model, and dynamically adjusting the power of the heating module; the intelligent clamping module recognizes the diameter of the tail and automatically adapts to and fixes the tail and the body of the mouse; the whole-body heat preservation module is used for maintaining mouse body temperature; and the energy storage power supply module is used for storing and supplying electric energy. By self-adaptive environment compensation and heating power adjustment, accurate temperature control is realized, experimental efficiency and data reliability are remarkably improved, animal stress response is reduced, and the experimental device is suitable for experimental requirements of tail vein injection.