48 results about "Tyrosine recombinase" patented technology

The invention discloses a construction method and application of a mouse model for specifically tracing heart valve cells, and belongs to the technical field of animal model construction. The model construction method comprises the following steps: (1) constructing a Wid1-Cre hybrid mouse before inserting a Cre recombinase coding gene into a termination codon of a mouse Wid1 gene; and (2) hybridizing the Wid1-Cre hybrid mouse with the Rosa26-tdTomato homozygous report mouse, so as to obtain the double transgenic mouse which simultaneously carries the Wid1-Cre gene and the Rosa26-tdTomato gene. According to the invention, on the basis of the expression characteristic that Wif1 is only limited to a valve area in the heart, Wif1-Cre is constructed; according to the Rosa26-tdTomato mouse model, specific tracing of heart valve cells is achieved, and an important experimental tool is provided for mechanism research of heart valve diseases and development of therapeutic drugs.

The invention discloses a system for controllably inducing pyroptosis of tumor cells and application of the system. The system comprises a red light activated Cre recombinase system and an expression vector of a GSDMDNT gene sequence of loxP flanking; the Cre recombinase system comprises: (1) an expression vector 1 containing a photosensitive protein PhyA gene sequence and a CreN59 sequence, and a connecting peptide located between the PhyA gene sequence and the CreN59 sequence; (2) an expression vector 2 containing a nucleoplasm shuttle protein FHY1 gene sequence and a CreC60 sequence, and a connecting peptide located between the FHY1 gene sequence and the CreC60 sequence; and (3) phycocyanobilin is contained. The system disclosed by the invention can be used for artificially and controllably regulating and controlling the expression of the cytotoxic gene GSDMDNT, so that directional and controllable pyroptosis induction is realized. The method can be applied to the fields of tumor therapy, gene therapy, cell biology and the like, and particularly has a wide application prospect in application scenarios requiring accurate control of gene expression.

The invention provides a construction method and application of a gene Idh1 conditional knockout mouse model and a preeclampsia heart injury animal model, and relates to the technical field of biology. According to the preeclampsia heart injury animal model, flox mice with conditional gene Idh1 knockout and mice with placental trophoblast specific expression Cre recombinase comate, PE-related phenotypes and typical symptoms of heart injury spontaneously appear in the double-positive pregnant mouse model, and the double-positive pregnant mouse model can be used for research of disease causes, diagnosis methods and drug research and development.

The present disclosure relates to enzymes, compositions and methods for performing conditional homologous recombination of a targeted DNA molecule or genome by using modified proteins comprising nucleic acid DNA binding proteins with protease-conditional recombinase (hereinafter “ProRec”) activity. Namely a new cre recombinase pro enzyme, compositions / kit and sensors comprising the cre recombinase pro enzyme of the present disclosure. Namely, a cre recombinase pro-enzyme, for identifying and quantifying proteolytic activity in a disease or an infection, comprising intein mediated circularization, a flip-excision cassette comprising an output reporter and / or effector protein; a linker sequence cleavable by a protease.

The invention discloses a rigorous induction system for corynebacterium glutamicum and application of the rigorous induction system, and belongs to the fields of genetic engineering and microbiology. The system can be strictly regulated when an inducer is not added, and can express only after the inducer is added. More specifically, the invention relates to the reduction of leak expression of Cre recombinase by comprising a logic gate, repression regulation and RNA regulation elements, and the use thereof for mediating SCRaMbLE to produce large-scale genome random rearrangement in Corynebacterium glutamicum synthetic genomes. Particularly, the method is used for reducing the leakage expression level of Cre recombinase and keeping the moderate expression level after induction. More importantly, the rigorous induction system disclosed by the invention can be applied to genome rearrangement to obtain rearranged strains with highly diversified genotype sequences, so that the purpose of increasing the yield of target products or enhancing the stress resistance of the strains is achieved.

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.

The purpose of the present invention is to provide a novel method for targeting a neuron population that is a neural circuit element which has more limited functional localization. The purpose of the present invention is also to provide a method for treating a disease or the like by targeting a specific neuron population involved in a disease or the like by said novel method and controlling the neural activity thereof.　The present invention is a vector set used for targeting a specific neuron population in a neural circuit, said vector set being characterized by comprising first to third vectors, wherein: the first vector is an antegrade trans-synaptic virus vector that contains a base sequence which codes for a split-Cre recombinase consisting of a C-terminal-side sequence of a Cre recombinase; the second vector is a retrograde trans-synaptic virus vector that contains a base sequence which codes for a split-Cre recombinase consisting of an N-terminal-side sequence of a Cre recombinase; the third vector is a non-trans-synaptic virus vector that contains an inverted sequence of a gene sequence which codes for a desired protein and that has a loxP sequence and a loxP mutation sequence which face opposite to each other and which are at both ends of the inverted sequence; and a functional Cre recombinase is formed in a cell that has been infected with the first vector and the second vector.

The present invention discloses a Cre recombinase regulatory system and its application, wherein the Cre recombinase regulatory system includes the ER50 gene. The present invention constructs a Cre recombinase regulatory system by introducing 4OHT to induce degradation of the protein ER50. In the Cre recombinase regulatory system constructed by the present invention, 4OHT achieves more stringent regulation of the functional gene Cre through dual regulation of ER50 and ERT2, reducing background issues in the Cre system. The application of this system will enable more rigorous spatiotemporal regulation of gene expression in cells and animals in the future, providing a more reliable research tool for conditional gene knockout in cells and animals.

This invention discloses a novel site-specific recombinase-mediated multisite chromosome editing tool and its applications. The invention provides a complete product suite for integrating or continuously integrating the genome of a host cell, comprising: a circular vector 1 composed of the following elements: loxF1, nox, ori, resistance gene, lox71, P3, P1, P2; and / or a circular vector 2 composed of the following elements: lox66, nox, ori, resistance gene, loxF2, P3, P1, P2; and a circular vector 3 for expressing Nigri and Cre recombinases; and a site-specific knock-in tool for knocking in the lox66-nox fragment at the desired integration site in the host cell genome; P1 and P2 are adapters, used to insert the DNA fragment to be integrated, and P3 is a universal primer. This invention enables rapid and continuous genome integration using simple molecular biology operations.

The invention discloses a construction method of an endothelial cell knockout Cdc42 gene aggravated bleomycin induced pulmonary fibrosis mouse model, which comprises the following steps of: mating a Cdc42 gene conditional knockout mouse with a mouse (Tie2-CreER) of which the endothelial cell specifically expresses tamoxifen induced Cre recombinase to obtain a double-transgenic mouse (Cdc42fl / fl-Tie2-CreER); the Cdc42 gene is specifically knocked out in vascular endothelial cells under the induction of tamoxifen, and then pulmonary fibrosis is induced by subcutaneous injection of bleomycin. The model shows aggravated pulmonary fibrosis phenotypes, including collagen deposition increase, alveolar structure damage and fibrosis-related protein expression up-regulation, and is high in construction success rate and good in repeatability. The invention also provides an application of the model in research of systemic sclerosis related interstitial lung disease pathogenesis and screening of anti-pulmonary fibrosis drugs, and an application of the Cdc42 gene as a drug target, and provides an accurate and reliable tool for pulmonary fibrosis research.

The invention discloses a construction method and application of a skeletal muscle muscle fiber conditionally induced knockout hnRNPK gene mouse model, and belongs to the technical field of animal models.The construction method comprises the following steps that 1, an hnRNPKflox / flox mouse and an ACTA1-CreEsr1 positive mouse are hybridized, and hnRNPKflox / +-ACTA1-CreEsr1 positive filial generation mice are screened out; (2) hybridizing the hnRNPKflox / +-ACTA1-CreEsr1 positive offspring mice with the hnRNPKflox / flox mice, and screening out the hnRNPKflox / flox-ACTA1-CreEsr1 positive mice, so as to obtain the hnRNPKflox / flox-ACTA1-CreEsr1 positive offspring mice, and screening out the hnRNPKflox / flox-ACTA1-CreEsr1 positive offspring mice, and (3) carrying out Tamoxifen treatment on the hnRNPKflox / flox-ACTA1-CreEsr1 positive mouse, activating Cre recombinase, cutting off the third intron fragment to the seventh intron fragment of the hnRNPK gene, and realizing selective knockout of the hnRNPK gene, namely constructing the mouse model for conditionally inducing and knocking out the hnRNPK gene from the skeletal muscle fiber. According to the invention, the vacancy in preparation of mice with skeletal muscle muscle fiber induced knockout hnRNPK genes in the prior art is filled, and a credible animal model is provided for revealing the functional research of the RNA binding protein hnRNPK in skeletal muscle growth and development regulation at an individual level.

The present application provides a method for directional transfer of heterologous chromosomes, which comprises the following steps: artificially adding a functional intron containing a loxP sequence into a drug resistance gene, splitting the drug resistance gene into two parts with an overlapping region containing a loxP sequence, inserting the two parts of the drug resistance gene into target chromosomes of a donor species and a recipient species respectively (the two parts of the drug resistance gene undergo Cre / loxP site-specific recombination under the action of a Cre recombinase, and the drug resistance function is recovered), and realizing directional transfer of heterologous chromosomes by means of heterologous cell fusion. The method can be widely applied to gene function research, construction of human disease models, development of therapeutic antibodies, and pharmacological and pharmacodynamic tests.

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of an exogenous gene cluster chromosome integration system in streptomyces. The invention reports a new way of integrating an exogenous gene cluster onto a streptomyces chromosome, namely RMCE. The RMCE reduces the influence of other DNA sequences on the stable existence of the gene cluster as much as possible; secondly, the RMCE landing site still has functions after recombination and can be reused; in addition, lox71 and lox66 sites are added on a Dre, Vika and PhiBT1 mediated integration vector for knockout of the resistance gene of the apolamycin, so that the resistance gene can be conveniently used for multiple times. In addition, the recombinase Cre, the recombinase Dre and the recombinase Vika are used for integrating gene clusters in the streptomycete for the first time, the function of the tyrosine recombinase in the streptomycete is expanded, and therefore the tyrosine recombinase has good practical application value.

The present invention provides a method for preparing a refractory epilepsy animal model and its application. Specifically, the present invention provides a method for preparing a refractory epilepsy animal model of a non-human mammal, wherein the preparation method comprises the following steps: (1) providing a non-human mammal A and a non-human mammal B expressing neuron cell-specific Cre recombinase of the same species; wherein the genome of the non-human mammal A has: (E1) an endogenous Cdkl5 gene, and (E2) a conditional knockout element operably linked to the Cdkl5 gene for conditional knockout of the Cdkl5 gene, wherein, in the presence of the Cre recombinase, the conditional knockout element conditionally knocks out the Cdkl5 gene in the genome of neuron cells, thereby inactivating the Cdkl5 gene; (2) mating and breeding the animal A with the animal B to obtain a progeny non-human mammal C with neuron cell-specific knockout of the Cdkl5 gene; (3) culturing the progeny non-human mammal C to obtain the refractory epilepsy animal model. The present invention can also preliminarily evaluate the efficacy of anti-epileptic drugs according to the changes in the spontaneous epilepsy phenotype of the animals.

Described herein is the discovery that TDP-43 proteinopathies may be induced in adult or neonatal animals bearing on one chromosome a mutant TARDBP gene encoding a mutant TDP-43 protein that lacks a functional nuclear localization signal (NLS) or a mutant TARDBP gene encoding a mutant TDP-43 protein that lacks a prion-like domain (PLD) and on the other homologous chromosome a TARDBP gene comprising a conditional knockout mutation. Knockout of the TARDBP gene comprising the conditional knockout mutation, e.g., using Cre recombinase, during the neonatal stage, e.g., at P0-P10, or during adulthood, e.g., at about 5 months of age, results in the mice exhibiting neuromuscular phenotypes such as early lethality, paralysis, weight loss, etc. These animals exhibit hallmark symptoms of ALS and that may be used in testing candidate agents useful in treating TDP-43 proteinopathies.

The present invention belongs to the technical field of animal model construction, and particularly relates to a method for constructing a spontaneous endometrial cancer mouse model and its application. The construction method includes constructing a mouse model with the genotype Cre; PTEN<supgt;flox / ‑< / supgt>; STK11<supgt;flox / flox< / supgt> through the Cre / loxP system. After activating the expression of Cre recombinase in uterine epithelial cells of this mouse model, the PTEN gene and the STK11 gene are selectively knocked out, thereby spontaneously inducing the occurrence of endometrial cancer. By selectively deleting the PTEN gene and the STK11 gene in uterine epithelial cells, the present invention rapidly changes the uterine pathology, and has the advantages of short tumorigenesis time and high tumorigenesis rate. Moreover, due to the use of in-situ tumorigenesis technology, the mouse model of the present invention can simulate the growth environment of human endometrial cancer, and the histological characteristics of the tumor are extremely close to those of human endometrial cancer, meeting the biological characteristics of human endometrial cancer.

The invention relates to the technical field of animal models, and particularly discloses a construction method and application of a mouse model with fat dystrophy complicated with atherosclerosis, and the construction method comprises the following steps: mating MED1flox / flox mice with Adipoq-CreERT2 mice to obtain F1-generation mice; the F1-generation mice and MED1flox / flox mice are subjected to backcross, the mice with the genotype being MED1fl / flAdipoq-Cre < + / -> are screened, tamoxifen is injected to induce fat cell specific Cre recombinase expression, and fat cell specific MED1 gene knockout mice are obtained; and performing caudal vein injection of AAV8-PCSK9 virus and feeding with a high-cholesterol and high-fat feed to obtain the fat dystrophy complicated with atherosclerosis mouse model. According to the method provided by the invention, the mouse model with fat dystrophy complicated with atherosclerosis is successfully constructed and obtained, and collaborative simulation and regulation of phenotypes of two diseases are realized.

The invention belongs to the technical field of animal model construction, and discloses a construction method of a drug-resistant depression animal model. A Cx43flox / flox mouse is constructed through a CRISPR / Cas9 technology, an AAV virus vector of Cre recombinase encoding a GfaABC1D promoter is further injected into the forehead cortex in a positioned mode, and a model mouse of the forehead cortex with the astrocyte Cx43 being conditionally knocked out is constructed. Through behavioral experiments and inflammatory reaction detection, depression-like behaviors of animals in the model and inflammatory reaction of the forehead cortex are evaluated, and treatment failure of antidepressant drugs on the model is observed. According to the invention, the drug-resistant depression animal model can be stably, simply and repeatedly constructed, and a research basis is provided for research on the pathogenesis of refractory depression and development of antidepressant drugs.

The application discloses a stringent induction system for corynebacterium glutamicum and application thereof, and belongs to the fields of genetic engineering and microbiology. The system can be strictly regulated without an inducer and can be expressed after the inducer is added. More specifically, it relates to reducing the leakage expression of Cre recombinase by containing logic gates, repressor regulation, and RNA regulatory elements, and its use for mediating the production of large-scale genomic random rearrangement of SCRaMbLE in the synthetic genome of corynebacterium glutamicum. In particular, it is used for reducing the leakage expression level of Cre recombinase and maintaining a moderate expression level after induction. More importantly, the stringent induction system of the application can be applied to obtain rearranged strains with highly diverse genotypic sequences in genomic rearrangement, so as to achieve the purposes of improving the yield of target products or enhancing the stress resistance of strains.

The invention discloses a construction method of a microglial cell conditional knock-down dynactin animal model, which comprises the following steps: selecting a Dctn1LoxP gene knock-in mouse, hybridizing the Dctn1LoxP gene knock-in mouse with a Cx3cr1-Cre transgenic mouse, and carrying out three rounds of breeding to obtain a genotype Dctn1LoxP / LoxP; a mouse of Cx3cr1-Cre; the expression of Cre recombinase is driven by a Cx3cr1 gene promoter, and the 2nd to 4th exons of the Dctn1 gene are cut, so that the knockout of p150Glue and the reduction of the protein levels of other subunits of dynactin are realized. The model is strong in specificity and high in knock-down efficiency, animals show microglial cell morphology abnormity and movement coordination function defects, the model can be used for researching the influence of dynactin on microglial cell neurobiological functions, and a reliable tool is provided for pathogenesis exploration and drug screening of related neurological dysfunction.

The invention relates to application of an Nlrp3 gene and / or a knockout reagent thereof in construction of an arterial vascular calcification animal model. The construction method of the arterial vascular calcification animal model comprises the following steps: (1) constructing a gene editing system for targeted knockout of a mouse Nlrp3 gene; (2) performing gene editing on a mouse fertilized egg by using the gene editing system in the step (1), and inserting loxP sites at two ends of the Nlrp3 gene; and (3) carrying out mating propagation on the mouse obtained in the step (2) and a mouse expressing Cre recombinase to obtain a mouse of which the Nlrp3 gene is knocked out, and culturing to obtain the arterial vascular calcification animal model. The arterial vascular calcification mouse is obtained by specifically knocking out the Nlrp3 gene of the mouse, and the arterial vascular calcification mouse has important application value in the aspects of mechanism exploration of arterial vascular calcification diseases and drug research and development.

The invention discloses a gene cluster cloning method and application thereof. The gene cluster cloning method comprises the following steps: performing restriction enzyme digestion on a target microbial genome to obtain a DNA fragment containing a target biosynthetic gene cluster; by utilizing a Gibson assembly method, connecting the upstream of the DNA fragment containing the target biosynthetic gene cluster with a p15Aorii-loxP-AmpR element containing pBE44 and a fragment aiming at a homologous arm of the upstream, and connecting the downstream of the DNA fragment containing the target biosynthetic gene cluster with a fragment containing a lacZ alpha-loxP-phage # imgabs0 # attP-traJ-ApmR element aiming at a homologous arm of the downstream and pBE45, so as to obtain a linear recombinant fragment formed by connecting three fragments; and electrically transferring the linear recombinant fragment into an escherichia coli host strain containing a Cre recombinase expression system, and cyclizing a large-fragment gene cluster in vivo by virtue of a Cre-loxP system, so as to obtain the annular recombinant plasmid. The method has the remarkable advantages of being easy and convenient to operate, high in cloning efficiency, high in universality and the like, and complex natural product gene clusters can be efficiently obtained.

The invention relates to the technical field of animal model construction, in particular to a construction method and application of a gene editing mouse model for studying calcium library function and low fertility in mature sperms.The model is characterized in that tamoxifen is used for inducing, a Ddx4ERT2 promoter is used for driving Cre recombinase expression, three subtypes of IP3R are specifically knocked out in sperms at the same time, and the mouse model is obtained. The method is used for researching the action and mechanism of the subtype in male fertility. After the model mouse is subjected to two rounds of induced knockout, the fertility is severely damaged, the sperm quantity and movement level are reduced, and the acrosome reaction is abnormal, so that the model is a good model for evaluating the sperm dysfunction, and is expected to be used for screening treatment drugs for improving the sperm dysfunction, especially male sterility diseases caused by advanced acrosome reaction in the future. In addition, through screening optimization of an IP3R agonist and a calcium signal detection platform, it is further confirmed that the mouse model is a good model for studying sperm calcium library mediated calcium signals.

The invention discloses a construction method and application of a lung cancer bone metastasis non-human animal model. The construction method comprises the following steps: step 1, constructing a Rosa26 fixed-point human-derived SLC7A7 conditional overexpression mouse model; 2, the R26LSL-slc7a7 homozygous mouse and KrasLSL-G12D / + are subjected to a reaction, and the mouse is subjected to a reaction; the method comprises the following steps: carrying out mating on a Tp53flox / flox mouse, carrying out multiple times of selfing on the obtained heterozygote to obtain a KPS mouse, and delivering an effective dose of Cre recombinase to the lung of the mouse. The invention successfully constructs a genetic engineering animal model capable of inducing three-gene collaborative manipulation, the model can efficiently and stably simulate the whole process of lung adenocarcinoma generation, development and distal bone metastasis, and provides a foundation for deeply exploring the pathogenic mechanism of lung cancer bone metastasis, exploring novel diagnostic markers and developing efficient anti-tumor drugs. And a stable and reliable experimental tool with good clinical correlation is provided.

The invention relates to the technical field of biological medicine, and discloses a method for constructing a Myh6-CRE transgenic golden hamster model and application of the Myh6-CRE transgenic golden hamster model. The invention provides a specific DNA (deoxyribonucleic acid) molecule. The specific DNA molecule sequentially comprises an upstream insulator, a Myh6 promoter, a Cre recombinase coding gene, a beta-globin polyA and a downstream insulator from upstream to downstream. The invention also provides a method for preparing the transgenic golden hamster, which comprises the following steps: introducing the expression construct containing the specific DNA molecule into a golden hamster fertilized egg in a fertilized egg microinjection manner, and randomly integrating the expression construct into a golden hamster genome, thereby obtaining the transgenic golden hamster. The invention further provides a Myh6-CRE transgenic golden hamster model, the transgenic golden hamster expresses Cre recombinase under the driving of a Myh6 promoter, and specific recombination between LoxP sites can be mediated. The Myh6-CRE golden hamster constructed by the invention can be used for establishing a conditional gene knockout model, and is suitable for researching the function and action mechanism of the Myh6 related gene in the occurrence and development process of heart and related diseases.

The present invention relates to the field of genetic engineering and synthetic biology, more particularly to means and methods for promoting genetic recombination. The present application discloses novel Cre recombinase dependent recombination sites that support methods of simultaneous cloning and testing.

The application discloses a screening marker recovery technology based on a Cre-LoxP combined Flp-Frt recombination system and application. The Cre-Loxp recombination system comprises a Cre recombinase and a LoxP site, and the LoxP nucleic acid combination comprises LoxP71 and LoxP66 sequences which can be specifically recognized and recombined by the Cre recombinase; similarly, the Flp-Frt recombination system comprises a Flp recombinase and a Frt site, and the Frt site comprises a Frt32 sequence which can be specifically recognized and recombined by the Cre recombinase. Due to the lack of screening markers and inducible promoters in the Schizochytrium, the site-specific recombinase cannot be recovered after being integrated into the genome, and therefore, the application aims to combine the two specific recombinase systems, so that the screening markers and the site-specific recombinase can be recovered unlimited times, and thus, the multi-gene editing of the Schizochytrium can be realized.

The present invention discloses a method and application for constructing an IL-11 knock-in mouse model. The present invention provides a specific DNA molecule, which comprises the following elements from upstream to downstream: an upstream homology arm, an mTFP1 encoding gene, a P2A self-cleavage peptide encoding gene, a Cre recombinase encoding gene, WPRE-polyA and a downstream homology arm. The present invention also provides a method for preparing a recombinant mouse, comprising the following steps: replacing the target segment in the mouse genomic DNA with the specific DNA molecule to obtain a recombinant mouse. The present invention constructs an IL-11-Cre tool mouse based on the Cre-LoxP recombination system, which can be used to solve the scientific problem of the IL-11+ regulatory molecular mechanism during the occurrence and progression of important diseases. The present invention will help to further explore the mechanism of action of IL-11 in the occurrence and development of diseases in the future, so as to find new therapeutic targets and develop more effective treatment strategies.

The invention provides a male testicular dysplasia mouse model as well as a construction method and application thereof, and belongs to the field of animal model construction. The construction method comprises the following steps: S1, inserting two loxP sites in the upstream and downstream of a mouse Dhx37 gene, and simultaneously expressing Cre recombinase to produce an F0-generation heterozygote mouse; s2, the F0-generation heterozygote mice and the wild mice are mutually mated to generate F1-generation heterozygote mice; then, the F1-generation heterozygote mice are mutually mated to generate F2-generation homozygote mice; s3, the F2 generation homozygote mouse and the whole body / tissue specific tool mouse are mated with each other to generate an F3 generation heterozygote Amh-Cre mouse; and then the F3 generation heterozygote Amh-Cre mice are mated with each other to generate an F4 generation homozygote Amh-Cre mouse. Male testis dysplasia mice can be massively produced through gene knockout and amplification breeding, and the stable character can be used as a basis for testis development research and drug screening.

The present invention discloses a recombinant adeno-associated virus composition and its application in precise sparse labeling. The adeno-associated virus composition comprises rAAV13 and rAAV9. The rAAV13 expresses Cre recombinase, which has the characteristic of limited infection. The rAAV9 expresses a fluorescent reporter gene dependent on the Cre recombinase. In vivo tests show that through the dual virus strategy, rAAV13 can be combined with rAAV9 after different multiple dilutions to achieve precise sparse highlight labeling. The present invention uses rAAV13 for precise sparse labeling for the first time, providing new tools and technical support for neuroscience research, and has broad application value and market prospects.