332 results about "Mus spretus" patented technology

The invention discloses a method for constructing a Ddx58-R109C point mutation mouse model by using a CRISPR / Cas9 gene editing technology and an application of the Ddx58-R109C point mutation mouse model. The method comprises the following steps: (1) target construction: searching a CDS region of a Ddx58 gene, determining an exon part, and determining a gene mutation site; (2) obtaining a fertilized egg, and injecting Cas9mRNA, gRNA and donor oligo into the fertilized egg of the C57BL / 6J mouse together in vitro through a microinjection technology to obtain an F0-generation hybrid mouse; (3) mating the F0-generation hybrid mouse with the wild type mouse to obtain an F1-generation mouse; (4) the F1-generation heterozygous mice which are identified to be positive are subjected to mating, and homozygous Ddx58-R109C mutant type and wild type mice can be obtained by breeding obtained offspring; (5) phenotypic analysis shows that Ddx58-R109C point mutation mice can have pathological changes of systemic lupus erythematosus and lupus nephritis; the invention can provide an effective research approach and method for researchers to deeply explore the action mechanism of Ddx58-R109C mutation in diseases, and has important significance for explaining the pathogenesis of systemic lupus erythematosus and researching and developing targeted therapeutic drugs.

The invention relates to a construction method of a macrophage specific fluorescence report mouse model, and belongs to the technical field of genetic engineering and genetic modification. According to the invention, a brand-new C1qc-CreChy gene knock-in mouse model is obtained after a C1qc gene locus is edited through a CRISPR / Cas9 genome editing technology for the first time, the expression of Cre and mCherry is directly driven by a C1qc promoter, and flow cytometry verifies that mCherry has very high expression quantity only in microglial cells in the brain of a mouse and macrophages in the spleen of the mouse. And mCherry expression is not detected in other immune cells, so that the two macrophages are traced and used for researching the functions of the macrophages. The invention provides a usable animal model for researching heterogeneity of macrophages, and provides a new animal model for researching complex immune functions in a body.

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 research method for researching an influence mechanism of high-fat diet on old-age health by using a mouse model, which comprises the following steps: S1, constructing a high-fat diet-induced mouse model: from normal diet feeding of mice to an old mouse stage, dividing the mice into two groups, and respectively performing high-fat diet feeding and normal diet feeding; s2, detecting physiological and functional indexes of the mice in the high-fat diet feeding group and the normal diet feeding group; s3, microbial diversity analysis: performing 16S rRNA sequencing to analyze the community structure of the intestinal microbiota of the mouse; s4, carrying out non-targeted metabonomics analysis: S4.1, carrying out intestinal tract non-targeted metabonomics sequencing; s4.2, carrying out non-targeted metabonomics sequencing on the liver tissues; s5, carrying out transcriptomics analysis; s6, performing conjoint analysis: performing the conjoint analysis on every two results of transcriptomics analysis, non-targeted metabonomics analysis and microbial diversity analysis; and S7, evaluating the influence of the high-fat diet on the health of the old rats by using the data obtained through analysis.

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 invention provides a sober mouse head fixing instrument capable of being adjusted in multiple dimensions, which comprises a first adjusting assembly, a second adjusting assembly and a window patch, and is characterized in that the first adjusting assembly is arranged on a chassis and has a displacement degree of freedom in a first direction and a rotation degree of freedom in a second direction relative to the chassis; the second adjusting assembly is arranged on the first adjusting assembly and has the rotation freedom degree in the first direction and the rotation freedom degree in the third direction relative to the chassis. And the window patch is arranged on the second adjusting assembly and has a displacement freedom degree in a plane formed in the second direction and the third direction relative to the chassis. Through the arrangement of the first adjusting assembly, the second adjusting assembly and the window patch, the fixing instrument for fixing the part, needing to be imaged, of an animal and performing multi-dimensional adjustment to be matched with imaging is achieved, the movement of the animal cannot be completely restrained, and the purposes that the head of a sober mouse is fixed, and behavioral experiment requirements can be met are achieved.

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 relates to the field of biological immunity, and discloses an immunotoxicity analysis and evaluation method and system.The method comprises the steps that immune cells and non-immune cells are cultured, an immune microenvironment of immune cell tissue is constructed, and an in-vitro immune system model of a human body is constructed; inputting an allogenic material into the in-vitro immune system model, and detecting cellular immune parameters of the experimental in-vitro immune system model; the method comprises the following steps: extracting a mouse genome of a mouse, constructing a gRNA-Cas9 compound of a target immune gene, implanting the gRNA-Cas9 compound into a preset mouse fertilized egg, culturing the target gene fertilized egg into a target gene mouse, inputting an exogenous material into the body of the target gene mouse, and analyzing humoral immune parameters of a blood sample; detecting organ parameters of the immune organs, calculating organ coefficients of the immune organs, and analyzing immune organ states of the immune organs; and analyzing an immunotoxicity evaluation report of the allogenic material on the human immune system. According to the invention, the accuracy of immunotoxicity analysis can be improved.

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 utility model relates to the technical field of laboratory mouse feeding, and discloses a laboratory mouse heat preservation feeding box which comprises a feeding box body, a supporting plate is fixedly connected to the inner side wall of the feeding box body, a supporting column is fixedly connected to the top of the supporting plate, and a connecting frame is fixedly connected to the top of the supporting column. The breeding box comprises a breeding box body, a connecting frame is fixedly connected to the outer side of the breeding box body, two symmetrically-distributed rotating columns are rotationally connected to the interior of the connecting frame, rotating rods are fixedly connected to the outer sides of the rotating columns, the number of the rotating rods is four, every two adjacent rotating rods are rotationally connected with each other, and connecting rods are fixedly connected to the outer sides of the rotating columns close to the side wall of the breeding box body. The outer side of the connecting rod is rotationally connected with a connecting piece, and the top of the supporting plate is fixedly connected with an air cylinder. The heat preservation breeding box for the laboratory mice has the advantages that the breeding box does not need to be opened, the mice in the breeding box are fed regularly and quantitatively, and the temperature in the breeding box is kept stable.

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 present invention provides a knock-in mouse model of a new mutation pathogenic site C104G of the Anxa11 gene and a construction method thereof, the method comprising: obtaining an active sgRNA, the nucleotide sequence of the sgRNA is shown in SEQ ID NO.2; obtaining an active Cas9mRNA, the nucleotide sequence of the Cas9 is shown in SEQ ID NO.3; mixing the active sgRNA, Cas9mRNA and a targeting donor with a nucleotide sequence as shown in SEQ ID NO.1 and microinjecting them into mouse fertilized eggs to obtain F0 generation mice; selecting F0 generation positive mice to mate with wild-type mice to obtain F1 generation mice with stable genotypes; and then screening to obtain Anxa11 gene mutation knock-in model mice. The method has high gene editing efficiency.

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 invention belongs to a molecular biological technology, and discloses a construction method and an application of sgRNA of targeted Mett13 as well as a transgenic mouse for knocking down Mett13 protein expression at a fixed point. A coding gene of guide RNA of Mett13 is obtained through Crispick online tool design, a PiggyBAC transposase system and a fertilized egg injection mode are utilized, a target gene is randomly inserted into a mouse genome, and then a TGsgMett13 overexpression transgenic mouse model is obtained through genotype identification and screening. The TGsgMettl3 overexpression transgenic mouse model can express a guide gene of Mettl3 in a whole body tissue, the TGsgMettl3 overexpression transgenic mouse model and a tissue-specific Cas9 overexpression mouse are hybridized to knock down the expression quantity of Mettl3 in a specific tissue in a mouse body, and a new tool and thought are provided for researching the Mettl3 gene function.

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.