235 results about "Mus spretus" patented technology

The invention relates to a humanized gene genetically modified non-human animal, particularly a genetically modified rodent, especially a genetically modified mouse, and in particular relates to a construction method of a humanized PD-L1 gene animal model and application of the model in the biomedicine field.

The invention discloses a sgRNA capable of specifically targeting Sirpa gene, a targeting vector, and a SIRPA gene humanized animal model. The invention also discloses a preparation method of an sgRNAvector of the humanized animal model, a preparation method of the humanized animal model and related application. The invention also relates to humanized genetically-engineered non-human animals, inparticular relates to genetically engineered rodents, in particular relates to genetically engineered mice, and in particular relates to a method for constructing the humanized SIRPA gene animal modeland use of the humanized SIRPA gene animal model in the field of biomedicine.

The invention relates to humanized gene modification of non-human animals, especially gene modification of rodents, and particularly gene modification of mice, and specifically relates to an establishment method of a humanized CD27 gene animal model and application thereof in the field of biomedicine.

The invention provides a construction method of a mouse model realizing conditional site-specific double-overexpression of HPV E6/E7 genes. The construction method comprises the following steps: 1, acquiring mice: (1) respectively designing and acquiring sgRNAs; (2) respectively preparing Cas9/sgRNA mixtures; (3) constructing a targeting vector; (4) carrying out microinjection and acquiring F0-generation mice, including F0-generation mice with HPV E6 knocked in at the specific ROSA26 site and F0-generation mice with HPVE7 knocked in at the specific H11 site; (5) acquiring F1-generation mice; and (6) acquiring F2-generation positive homozygote mice; and 2, acquiring conditional site-specific HPV E6/E7 double-knock-in model mice: hybridizing the two F2-generation positive homozygote mice toobtain the conditional site-specific HPV E6/E7 double-knock-in model mice. According to the method provided by the invention, the animal model which can be stably inherited and accurately regulated and controlled can be obtained.

The invention provides a construction method and an application of an HBeAg transgenic mouse model. The method comprises the following steps: CRISPR/Cas9 technology is adopted, an HBeAg gene is inserted into a pliver-HBeAg expression cassette at the Rosa26 gene locus in a fixed-point manner through homologous recombination, and a recombinant R26-e(Alb-HBeAg)1 targeting vector is constructed with an In-Fusion cloning method and contains a 3.3kb 5' homologous arm, pliver-HBeAg and a 3.3kb 3' homologous arm; Cas9mRNA, gRNA and the recombinant R26-e(Alb-HBeAg)1 targeting vector are micro-injectedinto a zygote of a C57BL/6J mouse, then the zygote is transplanted into the uterus of a C57BL/6J female mouse, and an HBeAg transgenic mouse is obtained through propagation and purification step by step. The transgenic mouse can specifically express HBeAg in liver, thereby being capable of applying to experimental animal models for researching HBeAg infection mechanism and evaluating treatment effects of therapeutic drugs and vaccines for chronic hepatitis B.

The invention discloses a PirB gene-knock-in mouse animal model, which comprises gRNA1 and gRNA2 for determining specific target sites to be knocked in by PirB gene, wherein the PirB gene is knocked into an intron 1 of ROSA26 gene of a C57BL/6J mouse; the gene sequence of the gRNA1 is shown in SEQ ID NO.1; and the gene sequence of the gRNA2 is shown in SEQ ID NO.2. The invention further specifically discloses a construction method of the mouse animal model, the mouse animal model provides a good foundation for research on PirB gene function and in-vivo verification. Hybridization of PirB-knock-in animal model with different types of Cre mice can be used to study the functions of PirB in different organs or different cell types or different disease models.

The invention provides a kind of transgenic inhuman mammal by producing klone gene without mouse model. Roll outside self murder gene into the mammal. The expression box contains promoter, self murder gene relation with promoter and termination codon. The natural masculine gene will express in testes organize after transgenic animal builds system. The transgenic animal can provide blastosphere for eliminating gene technic to improve the efficiency of the technic and to solve the problem that mouse genetic background is impure.

The invention provides a construction method of a mouse model with conditional over-expression of HPV E7 gene at H11 locus. The construction method of the mouse model with the conditional over-expression of the HPV E7 gene at the H11 locus comprises the following steps: designing and obtaining sgRNA; (2) preparing a Cas9/sgRNA mixture; (3) constructing a targeting vector; (4) co-injecting the targeting vector and the Cas9/sgRNA mixture into fertilized eggs of a mouse so as to subsequently obtain F0 generation mice; and (5) obtaining F1 generation mice. The construction method of the mouse model with the conditional over-expression of the HPV E7 gene at the H11 locus solves the problem that "nude mice are not suitable for studying role of HPV E7 in HPV E7-mediated processes of tumor microenvironment immune regulation, immune escape and the like for the nude mice lack competent immune systems" in the prior art. The mouse model provided by the invention has a competent immune system, so that the mouse model is suitable for all researches on interaction between the HPV E7 and the immune system.

The invention discloses an Il21 gene knockout mouse model and a construction method and an application thereof. The construction method comprises the following steps that S1, based on the CRISPR/Cas9 technology, a knockout area of an Il21 gene is determined, specific target sites sgRNA1 and sgRNA2 are designed according to the determined knockout area, and gene sequences of the gRNA1 and the gRNA2 are shown in SEQ ID NO.1 and SEQ ID NO.2 respectively; S2, microinjecting active gRNA1, gRNA2 and cas9 proteins into fertilized eggs of the mouse, transplanting the fertilized eggs survived after injection into the body of the false pregnant female mouse, and obtaining an F0-generation mouse after the false pregnant female mouse is pregnant and farrowing; S3, mating the F0-generation mouse obtained in the step S2 with a wild-type mouse to obtain an F1-generation heterozygote; and S4, carrying out inbreeding on the F1-generation heterozygote obtained in the step S3 to obtain F2-generation mice, namely the IL21 gene knockout mouse model. Based on the CRISPR/Cas9 technology, the mouse animal model is established by knocking out an exon3-exon5 region in an ENSMUST000029273.7 transcript of an IL21 gene for the first time by designing reasonable gene sequences of specific target sites gRNA1 and gRNA2, and the mouse animal model can be applied to related research after tumor healing.

The invention discloses primers, probe, kit and method for the RPA-LFD visualization rapid detection of Schistosoma nucleic acid. Sequences of the primers are as shown in SEQ ID NO.1-2, and a sequenceof the probe is as shown in SEQ ID NO.3. The RPA primers and probe are designed with the SjCHGCS19 gene as a target sequence, and the sensitive and rapid visualization detection of the schistosome nucleic acid is realized by using a RPA amplification technology combined with lateral flow chromatography test strip method. The detection method of the invention has a detection limit of 1 fg for Schistosoma japonicum genome DNA, and is expected to be used in the general detection of Schistosoma mansoni and Schistosoma haematobium. The method can detect circulating nucleic acid of Schistosoma in the serum of mice at an early stage of infection, the operation is simple and fast, no special equipment is required, a reaction temperature is close to room temperature, results can be observed with the naked eye, and the realization of the early and sensitive and rapid detection of intermediate hosts of Schistosoma on site, and the timely and sensitive monitoring of environments with high schistosomiasis transmission risks are facilitated.

The invention discloses a construction method and application of a preeclampsia mouse animal model. According to the construction method, the m6A modification level in a mouse is reduced, and a mutant mouse with the reduced RNA m6A modification level, namely the preeclampsia mouse animal model, is obtained; specifically, a Mett13<+/>hybrid mutant mouse is constructed through an ES targeting technology, and a result shows that the sensitivity of the mutant mouse to a PE inducer L-NAME is increased, the mutant mouse presents the characteristics of PE, and a new thought and a new means are provided for etiological diagnosis, prevention and treatment of preeclampsia.

The invention discloses a urine biomarker for Japanese schistosomiasis early diagnosis. The urine biomarker is one or more of a xanthurenic acid, a naphthalenesulfonic acid, or enanthylcarnitine. Sensitivity and specificity of the marker are greater than 0.9, and an area under the curve (AUC) is greater than 0.9, indicating that the three indicators have better predictability and may be used for the Japanese schistosomiasis early diagnosis. The invention further discloses a metabolomics-based screening method for the urine biomarker for the Japanese schistosomiasis early diagnosis. By constructing a mouse Japanese schistosomiasis model, ultra performance liquid chromatography-tandem mass spectrometry is used to perform metabolomics analysis on mouse urine, and a characteristic differentialmetabolite, that is, the biomarker for the Japanese schistosomiasis early diagnosis, between a normal mouse and a Japanese schistosomiasis-infected mouse is found and analyzed. The screening method is non-invasive, convenient and fast, and can accurately reflect a metabolic spectrum difference between the Japanese schistosomiasis-infected mouse and the normal mouse and have high specificity.

The invention provides a construction method of an SDK2 gene mutation mice model. The mice model comprises the following steps of designing and constructing sgRNA capable of specifically identifying an SDK2 gene on the basis of a CRISPR/Cas9 system, injecting the sgRNA and a targeting vector constructed on the basis of the sgRNA into a mice fertilized egg; and after embryo transplantation, screening out F0-generation mice with SDK2 gene mutation from the output mice, and hybridizing the F0-generation mice with wild type mice to obtain an F1-generation mice model with SDK2 gene mutation. The mice model constructed by the method can be stably passaged, the action mechanism of the SDK2 gene in mice hereditary cataract can be conveniently researched in practical application, and under the condition that human patient research materials are not easy to obtain and are restricted by medical ethics, the SDK2 gene can be stably cloned. The mice model provided by the invention can become an important tool in the research of hereditary cataract, and a research model capable of realizing stable heredity is provided in the research of pathogenesis, treatment methods, drug screening, cataract surgery and the like.

The invention relates to the technical field of biology and in particular relates to a method for constructing a hepatitis B virus (HBV) infected mouse model and application of the HBV infected mouse model. The invention provides a method for constructing a high-ALT (alanine transaminase) HBV infected mouse model. The method for constructing the HBV infected mouse model comprises the step of medicating HBV genome DNA plasmid to a Tlr2 gene-deficient mouse, so that the HBV infected mouse model is constructed. The mouse model provided by the invention is relatively simple to construct, relatively easy in technological means and low in construction cost; besides, the adopted HBV plasmid vector is non-toxic, can not induce immune response of the mouse and also can avoid serious immunodeficiency in a chimera mouse model, and the obtained HBV infected model has higher blood serum ALT level and obvious liver inflammation expression.

The invention relates to a preparation method of a transgenic mouse of specificity expression Cre recombinase of a hematopoietic system. The invention discloses a transgenic mouse animal model, a deoxyribonucleic acid (DNA) fragment pEDAG-Cre is integrated in a genome of the transgenic mouse animal model, the DNA fragment comprises a promoter at an end of a gene (EDAG) 5' which is related to human embryonic development, a Cre recombinase gene and a human growth hormone gene polyA fragment. The invention also comprises a preparation method of the transgenic mouse animal model. The method comprises the steps of cloning 2.3 kb of a control region of the promoter at the end of the EDAG gene 5' by using a polymerase chain reaction (PCR) method, constructing a transgenic expression vector pEDAG-Cre, leading 5.6 kb of transgenic fragments into a male pronucleus of the mouse by using a micro-injection method, and identifying a positive transgenic mouse by using the PCR method. The RT-PCR method and a mouse-identifying result report ROSA26LacZ show that the specificity expression Cre recombinase of the transgenic mouse can be achieved only in the hematopoietic system.

The invention relates to mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging. The strain is conserved in the China General Microbiological Culture Collection Centre specified by the State Intellectual Property Office on March 13, 2012, and the conservation number is CGMCCNo.5893. The mouse typhus salmonella UF110 is a strain obtained by the fact that a spv gene on a virulence plasmid is knockout, and can be used for researching the function of mouse typhus salmonella virulence gene spv as a control. The UF110lux is a strain which is constructed on the basis of the mouse typhus salmonella UF110 and has the stable biological luminescence property. According to the invention, the shortage of a traditional fluorescent mark is overcome, and the invasion, proliferation and diffusion of the mouse typhus salmonella in a living body of mice can be monitored dynamically in real time by using a living body imaging instrument. The invention provides a convenient tool for further study on a pathogenic mechanism of mouse typhus salmonella.

The invention relates to a test sample allergenicity detection method and a kit thereof. The detection method comprises the following steps: randomly dividing a plurality of mice into groups; measuring the thickness of ears of each mouse of different groups; entering into a first time period; within the first time period, smear an equal amount of a reagent on the ears of each mouse of different groups; entering into a second time period; when the second time period is ended, firstly measuring the thicknesses of the ears of the mice of different groups, further killing the mice, taking down lymph glands after the ears of the mice under a sterile condition, preparing a cell suspension, putting a 5-bromodeoxyuridine solution into the cell suspension, culturing, and testing the content of the5-bromodeoxyuridine by using an ELISA (Enzyme-Linked Immuno Sorbent Assay) method; and judging the allergenicity of a test substance according to limiting standards. By adopting the method, whether the test substance has positive allergenicity can be tested, intraperitoneal injection is avoided, then animals can be prevented from pain, the amount of the 5-bromodeoxyuridine can be greatly reduced,and the detection cost can be reduced.

The invention discloses a method for constructing an Fzd6-Q152E site-directed mutagenesis mouse model based on CRISPR/Cas9 and application. It is found through comparison that the positions of rs61753730 in different species have certain conservative property. Therefore, the method comprises the steps of targeting positions of rs61753730, namely Q152E, in a mouse, designing andn synthesizing sgRNA, corresponding primers and Donor sequences, constructing an sgRNA/Cas9 expression vector, then performing in-vitro transcription after the vector is subject to linearization and purification, then mixing sgRNA, Cas9mRNA and Donor, putting the mixture into fertilized eggs of the mouse is through micro-injection, and then transplanting the survived fertilized eggs into the uterus of a pseudopregnant female mouse, wherein the born mouse is an edited F0-generation mouse; and mating a positive mouse with a wild mouse to obtain F1-generation mice through PCR and Sanger sequencing identification, finally mating F1-generation heterozygous mice which are identified to be positive, and breeding the obtained progeny to acquire a homozygous Fzd6-Q152E mutant and wild mouse. The model mouse can be used for researching the biological function of rs61753730.

The invention discloses a recombinant avian influenza virus carrying NanoLuc gene and application thereof in a living imaging mouse model. The invention provides a DNA fragment of which the nucleotidesequence is as shown in SEQ ID NO.1, and the DNA fragment can be used to construct a recombinant plasmid, a recombinant virus and a mouse model. In the invention, a recombinant avian influenza viruswhich is similar to the wild type virus in the replication ability and pathogenicity is constructed. In the SPF chick embryo passage, the NanoLuc luciferase gene with good stability can be detected inthe collected 4 generations of virus solutions. In the invention, a living imaging mouse model which has a visualization function is successfully constructed, and the distribution position of the influenza virus in a mouse can be detected without dissecting the mouse. Longitudinal assessment can be performed to observe the dynamic process of influenza virus infection in the same mouse for a continuous period of time, and a visualization tool is supplied for the study of influenza virus.

The invention discloses a postmortem interval deduction method based on microbiome sequencing data and machine learning algorithms. The method comprises the following steps: by taking DNA (deoxyribonucleic acid) which is extracted from cecum samples of dead bodies of mice as a template, designing primers for 16S rDNA of microorganisms, performing (polymerase chain reaction) amplification constructing a gene library by using amplification products, and performing sequencing; performing OTUs (operational taxonomic units) clustering and species taxonomy analysis on effective sequencing data; andon the basis of abundance distribution situations of species, removing species classes with small relative abundance and interclass variance variation, performing extraction so as to obtain characteristic microorganism varieties for constructing a postmortem interval deduction model, and training a artificial neural network by using sample microorganism abundance data at different time points after death of the mice, so as to obtain a mathematic model which is capable of accurately predicting postmortem intervals.

The invention provides a construction method of an animal model with a liver HO-1 gene conditionally knock out by utilizing CRISPR-Cas9, belonging to the technical field of gene engineering. The method comprises the following steps: 1) introducing an sgRNA sequence, a Cas9 gene sequence and a donor gene sequence into fertilized eggs of a mouse to obtain an F0-generation mouse containing a targeting sequence; 2) hybridizing the F0-generation mouse containing the targeting sequence obtained in the step 1) with a wild-type mouse to obtain a heterozygous mouse containing the targeting sequence; and 3) hybridizing the heterozygous mouse containing the targeting sequence obtained in the step 2) with an Alb-Cre positive mouse to obtain the liver HO-1 gene knockout animal model.

The invention discloses a serum biomarker for early diagnosis of schistosomiasis japonica. The serum biomarker is phosphatidylcholine and/or palmitoyl choline. The sensitivity and specificity of the marker are both greater than 0.9, and the under-curve area (AUC) is greater than 0.9, which indicates that the two indexes have favorable predictability and can be used for early diagnosis of schistosomiasis japonicum katsurada. The invention further discloses a screening method of the serum biomarker for early diagnosis of schistosomiasis japonica based on metabonomics. A mouse schistosomiasis japonica model is constructed, and the metabonomics analysis is carried out on mouse serum by utilizing an ultra-high performance liquid chromatography-tandem mass spectrometry technology, and characteristic differential metabolites between a normal mouse and a schistosomiasis japonica infected mouse are found and analyzed, namely, the schistosomiasis japonica early diagnosis biomarker is obtained. The screening method has the characteristics of convenience and quickness, can accurately reflect the metabolic spectrum difference between a schistosomiasis japonicum infected mouse and a normal mouse, and is high in specificity.