59 results about "Animal gene" patented technology

The invention discloses a Parkinson's disease animal model, a construction method and application, and relates to the field of animal genetic engineering and disease model construction. According to the model, seven mutation sites related to the human familial Parkinson's disease are introduced behind an initiation codon of a fourth exon of an endogenous SNCA gene, and the mutation sites are A18T, A29S, A30P, E46K, H50Q, G51D and A53T and have genotypes related to the human familial Parkinson's disease. The pig model shows nigra dopaminergic neuron reduction and cortical neuron reduction in the newborn period, and the core pathological process of the human Parkinson's disease can be systematically reproduced. The model can be widely applied to Parkinson's disease pathogenesis research, drug screening and cell therapy effect evaluation, and has important scientific research and preclinical application values.

The application belongs to the technical field of animal genetic engineering, and particularly discloses a construction method of C1R gene humanized animal cells and animal models and application thereof. The construction method of the C1R gene humanized animal cells comprises the following steps: introducing a human C1R gene into non-human animal cells, so that the human C1R gene is expressed in the non-human animal cells to produce humanized C1R protein, and meanwhile, the expression of an endogenous C1ra gene in the non-human animal cells is reduced or eliminated. The application preferably adopts CRISPR / Cas9 gene editing technology to accurately insert a genomic sequence encoding a human C1R mature protein into a 2nd exon region of a mouse C1ra gene, while retaining a mouse source promoter, a 5' untranslated region (UTR) and a signal peptide sequence. A mouse cell or model capable of specifically interacting with an anti-human C1R antibody is successfully constructed.

The invention is applicable to the technical field of animal genetic engineering, and provides a DHX36 gene molecular marker related to bovine sperm motility traits and application thereof, the DHX36 gene molecular marker is a 231st locus of a bovine DHX36 gene segment as shown in a sequence table SEQ ID NO: 1, and the locus has a C-G base mutation. Based on the DHX36 gene molecular marker, the genotype is judged through sequence determination, and the genotype can be associated with the normal and unfrozen activity traits of bovine sperms. An analysis result shows that the bovine sperms of individuals with different genotypes are normal and have significant differences in vitality after unfreezing. According to the invention, the specific primer group is utilized to obtain the bovine DHX36 gene segment related to the normal and unfrozen activity traits of bovine sperms, and the specific SNP site in the segment is taken as the molecular marker, so that a theoretical basis and specific application are provided for marker-assisted selection of cattle.

The invention relates to a gRNA of a targeted non-human animal Zmpste24 gene, a preparation method of a Zmpste24 gene knockout non-human animal model and a phenotype identification method. The gRNA comprises a nucleotide sequence which is complementary with a second exon region of the non-human animal Zmpste24 gene; the target sequence of the second exon region of the non-human animal Zmpste24 gene is as shown in SEQ ID NO: 4; the gRNA comprises gRNA1 as shown in SEQ ID NO: 1 and gRNA3 as shown in SEQ ID NO: 3, and the molar ratio of the gRNA1 to the gRNA3 is (1-5): (1-5). The Zmpste24 gene knockout non-human animal model prepared by the invention shows a remarkable senescence phenotype at the age of 12 weeks, and has the advantages of remarkable phenotype, good reproducibility, short experimental period and the like. The multi-dimensional phenotype identification method established by the invention can comprehensively evaluate the influence of gene knockout on the aging process.

The invention discloses a homologous recombination system, a transposition system, a fruit fly animal gene high-throughput screening method and application. The screening method comprises the following steps: firstly, separating and establishing a stable cell line from a fruit fly animal; then co-transfecting a CRISPR / Cas9 mediated homologous recombination system, and obtaining a cell line for stably expressing Cas9 protein through puromycin screening and a limited dilution method; the sgRNAs plasmid library constructed by combining a PiggyBac transposon system can realize efficient screening and functional analysis of a target gene in the Cas9 cell line. The high-throughput screening method provided by the invention is widely applicable to gene function research and target gene screening, is simple in preparation process and convenient to operate, has good universality, and provides effective technical support and solution for related fields.

This application discloses a smart method for preparing gene-edited mammalian cells. The method includes: acquiring the target gene sequence information of the mammal to be observed and processing it according to preset screening rules to obtain the target sgRNA sequence; mixing an RNP complex suspension prepared based on the target sgRNA sequence with a suspension of embryonic stem cells to be edited, and performing electroporation transfection using multiple consecutive micropulses to obtain a transfected cell suspension; acquiring raw images of the transfected cell suspension and preprocessing them to obtain single-cell images, and using a trained lightweight convolutional neural network model to obtain the predicted gene-editing efficiency and prediction confidence after 48 hours; if the transfected cell suspension meets the quality control conditions, expanding and culturing the transfected cell suspension to obtain the prepared gene-edited cells. This method can shorten the cell culture cycle, reduce the waste of cell materials, improve the stability of transfection efficiency, and achieve precise preparation of gene-edited cells.

The invention discloses a construction method of a mouse model capable of specifically photoactivating oligodendrocyte precursor cells, and belongs to the technical field of animal genetic engineering. On the basis of a Cre-loxP recombinase system, NG2 is selected as a specific promoter of oligodendrocyte precursor cells, an NG2-CreERT: cOpn5 mouse expressing photosensitive protein cOpn5 under specific driving of NG2 is constructed, and compared with a traditional method for injecting photosensitive protein cOpn5 through viruses, the mouse model can avoid tissue damage caused by the method; meanwhile, the method can also be used for functional research of the oligodendrocyte precursor cells, so that the research on the effect of the oligodendrocyte precursor cells in a nervous system is facilitated, and a new strategy and a new method can be possibly provided for treating related nervous system diseases and developing drugs of the related nervous system diseases.

The present application relates to a kind of gRNA targeted to non-human animal OPG gene, preparation method of OPG gene knockout non-human animal model.The preparation method of OPG gene knockout non-human animal model includes the following steps: using gene editing technology, destroy non-human animal OPG gene, for the gRNA targeted to non-human animal OPG gene includes one or two shown in SEQ ID NO:1 And / or SEQ ID NO:2.The present application designs the gRNA specifically targeted to non-human animal OPG gene, using Cas9 protein, OPG gene is knocked out, causes frame shift mutation, to reach the purpose of gene knockout to non-human animal.The method of the present application to non-human animal OPG gene modification is simple and easy to operate, short cycle, using the method of OPG model for research OPG gene physiological mechanism and molecular mechanism provides foundation, while providing important tool for research osteoporosis pathogenesis and drug development.

The invention discloses an NDUFS2 gene heterozygous editing pig as well as a construction method and application thereof, and relates to the field of animal gene engineering and disease model construction. The eighth exon of the NDUFS2 gene of the pig is deleted or mutated, and presents an NDUFS2 heterozygous genotype; in the newborn period, the number of neurons of the middle cerebral cortical layer and the number of dopaminergic neurons are obviously reduced, the number of neurons of the adult cortical layer is continuously reduced along with mitochondrial swelling and crest fracture, and after MPTP treatment, the number of neurons of the adult cortical layer is disordered in motion trail, and the body tremor index is increased. The pig model not only can be used for revealing the action mechanism of the NDUFS2 defect in neurodegenerative diseases (such as Parkinson's disease), but also can be used as an important tool for screening new drugs, evaluating treatment means and researching mitochondrial diseases.

The present invention belongs to the field of animal genetic engineering and genetic modification. Specifically, it discloses a method for constructing a humanized epidermolysis bullosa DEB mouse model and its application in biomedicine. The method comprises the following steps: preparing genetically engineered mice using mouse embryonic stem cells, and replacing the full-length mouse Col7a1 gene sequence with a human COL7A1 genomic sequence or a mutant sequence thereof; preferably, the human COL7A1 genomic sequence is represented by hg19:chr3:48,635,720-48,603,572; the human COL7A1 genomic mutant sequence is represented by hg19:chr3:48,635,720-48,603,572&c.6527 dup C; the mutant animals constructed by the present invention can survive for 7-10 days, which is significantly longer than the median survival of 2 days reported for Col7a1- / - knockout mice, without drug maintenance, providing a longer window of survival for therapeutic experiments.

Provided is a method for constructing a humanized FKBP8 gene knock-in animal model, comprising: (1) constructing a targeting vector for humanized FKBP8 gene knock-in, and transfecting the targeting vector, Cas9, and sgRNA into fertilized eggs of donor animals; (2) transplanting surviving fertilized egg cells into the oviduct of pseudopregnant animals to obtain F0 generation animals and identifying the F0 generation animals, wherein animals showing positive homologous recombination are chimeric animals; and (3) mating the chimeric animals with wild-type animals to obtain F1 generation animals and performing identification, wherein F1 animals showing positive homologous recombination are humanized FKBP8 knock-in animals. In the construction method, an FKBP8 gene sequence in a donor animal is substituted by a human FKBP8 gene sequence, so as to achieve the purpose of humanizing the FKBP8 gene in the donor animal, thus successfully establishing a humanized FKBP8 gene knock-in animal model, so that screening for drugs targeting the FKBP8 gene and a product thereof in the animal model, or studying the pathogenesis and treatment mechanisms of the FKBP8 gene and the product thereof in related clinical diseases can be realized.

Provided is a universal preclinical bio-distribution detection kit for NK cell therapy products. The present invention further relates to specific use of a gene sequence set forth in SEQ ID No. 1 in distinguishing a human NK cell gene from a non-human animal gene. By means of providing a specific DNA sequence, a DNA sequence derived from the human NK cell can be specifically distinguished from a DNA sequence derived from the non-human animal gene, and based on the specific DNA sequence, a qPCR system for distinguishing the human NK cell gene from the non-human animal gene is constructed, providing a primer pair and a probe set for NK cells and derivative cell therapy products thereof. In addition, a universal preclinical bio-distribution detection kit for the NK cell therapy products is successfully designed, thereby providing convenience for the preclinical research on the NK cells and derivative cell therapy products thereof.

The invention belongs to the technical field of animal gene breeding, and particularly discloses a functional mutation site combination based on a bovine immune disease-resistant tissue regulation region and application. According to the chip, the effect of bovine genome mutation sites on the influence of the activity of an immune tissue genome regulation element is fully considered, and 9394 mutation sites (SNP) which highly interfere with the binding strength of bovine transcription factors are screened from a large number of bovine genome mutation sites and immune tissue epigenetic regulation data; each marker provided by the invention is more closely associated with gene expression regulation and phenotype, and cattle disease resistance related character marker-assisted selection or cattle disease resistance breeding can be carried out by adopting a functional mutation site combination based on a cattle immune disease-resistant tissue regulation region.

This invention belongs to the field of animal genetic engineering technology, specifically relating to a cashmere goat circRNA, an interference vector, and their application in regulating cashmere goat hair follicle development. The cashmere goat circRNA is circ-0000987; the base sequence of circ-0000987 is shown in SEQ ID NO.1, and it is used to regulate cashmere goat hair follicle development. This invention reveals for the first time the inhibitory effect of circ-0000987 on the proliferation of cashmere goat papillary cells. The results of this invention show that circ-0000987 plays a proliferative inhibitory role in cashmere goat papillary cells, inhibiting their activity by negatively regulating cell proliferation-related pathways; and participating in the transition of hair follicles from the anagen to catagen phase by inhibiting cell proliferation and promoting apoptosis.

This invention discloses PCR primers, interspecific hybridization methods, and applications for identifying interspecific hybrid offspring of swimming crabs, belonging to the field of aquatic animal genetic engineering technology. The PCR primers of this invention include an upstream primer as shown in SEQ ID NO.1 and a downstream primer as shown in SEQ ID NO.2. When using these PCR primers for PCR amplification, a 786 bp band indicates *Portunus trituberculatus*; a 632 bp band indicates *Portunus cerana*; and a 963 bp band indicates *Portunus rubrum*. The hybrid offspring simultaneously exhibit two specific amplification bands, thus achieving rapid and accurate identification of the hybrid offspring. This invention establishes for the first time a method for identifying hybrid offspring based on PCR amplification fragment length polymorphism, which has advantages such as simple operation, low cost, high accuracy, and suitability for large-scale detection. It can be applied to the fields of swimming crab germplasm innovation, new variety breeding, and genetic improvement.

The invention discloses a CRHR1 gene molecular marker for detecting bovine superovulation traits and an application method, and belongs to the technical field of animal gene engineering, the length of a nucleotide sequence of an obtained CRHR1 gene segment is 666bp, a c-t base mutation exists at the 455bp position of the segment, the mutation site is used as a molecular marker, and the mutation site is used for detecting the superovulation traits of cattle. The genotype is judged through sequence determination, and the genotype can be associated with the excellent ovulation character of the cattle. An analysis result shows that the superovulation performance of individuals with different genotypes is remarkably different. According to the invention, a specific primer is utilized to obtain a CRHR1 gene partial segment related to the excellent superovulation character of cattle, and a specific SNP site in the segment is taken as a molecular marker, so that a theoretical basis and specific application are provided for marker-assisted selection of cattle.

The invention belongs to the technical field of animal genetic engineering and biological breeding, and particularly relates to a recombinant plasmid for co-expressing human antibacterial peptide and human interferon and application of the recombinant plasmid in the aspect of improving the disease resistance of aquarium fish. The recombinant plasmid is a plasmid which contains a human antibacterial peptide gene hLL-37 and a human interferon gene hIFNA1 and has a Tol2 transposon, a goldfish pan-expression promoter gActb1, a green fluorescent protein gene EGFP and a polyA sequence, and the nucleotide sequence of the recombinant plasmid is as shown in SEQ ID NO. 1. The recombinant plasmid and the Tol2 transposase mRNA are jointly introduced into the aquarium fish, so that the antiviral capability and the antibacterial capability of the aquarium fish can be improved.

The application belongs to the technical field of genetic engineering of aquatic animals, and discloses application of fish VDAC2 genes in resisting inflammation reaction induced by aquatic viruses. Experimental data of the application show that the inflammation reaction caused by SVCV can be significantly inhibited by using VDAC2 inhibitor Dids or knocking down VDAC2, and the survival rate of fish after infection with the SVCV virus can be effectively improved, so VDAC2 plays an important role in regulating the inflammation reaction induced by the spring viremia of carp virus. The application provides a new drug target for virus prevention and control in the field of aquatic products, and has important significance for prevention and control of aquatic viruses.

The invention discloses a mutant gene of a gm15262 gene and application of the mutant gene, and belongs to the technical field of molecular biology and medicine. The CDS sequence of the mutant gene of the gm15262 gene is shown as SEQ ID NO.1, and compared with the CDS sequence of a normal human source gm15262 gene, the CDS sequence of the mutant gene of the gm15262 gene is subjected to c.C338T (p.P113L) mutation. A male sterility animal model can be constructed by enabling corresponding sites of the animal gm15262 gene to be subjected to same mutation, when the animal model is a mouse model, a male sterility mouse model can be obtained by enabling the gm15262 gene of a mouse to be subjected to c.C305T (p.P102L) mutation, and the mouse model is used for accurately simulating pathogenic mutation of human genetic diseases. And a key experimental platform is provided for development of gene therapy, targeted drug screening and germ cell editing research. The invention has important significance in diagnosis and treatment of male infertility.

The invention is applicable to the technical field of animal genetic engineering, and provides an FETUB gene molecular marker related to bovine sperm motility traits and application thereof.The FETUB gene molecular marker is an SNP site on a bovine FETUB gene segment as shown in a sequence table SEQ ID NO: 1, specifically, the 315th site of the bovine FETUB gene segment, the site has an A-G base mutation, and the site has an A-G base mutation. The generation of gene single nucleotide polymorphism is caused. Based on the FETUB gene molecular marker, the genotype is judged through sequence determination, the genotype can be associated with the normal and unfrozen activity traits of the bovine sperms, and the normal and unfrozen activities of the individual bovine sperms with different genotypes have significant differences. The FETUB gene molecular marker provided by the invention provides a theoretical basis and specific application for marker-assisted selection of cattle.

This invention relates to the field of animal genetic engineering technology, providing a molecular marker for the ATP5PO gene associated with bovine sperm motility and its application. The ATP5PO gene molecular marker is located at position 203 of the bovine ATP5PO gene fragment, where an A-G base mutation occurs, resulting in a single nucleotide polymorphism (SNP). The nucleotide sequence of the bovine ATP5PO gene fragment is shown in SEQ ID NO:1. Based on this ATP5PO gene molecular marker, genotype can be determined through sequencing, and the genotype can be associated with normal and thawed bovine sperm motility. This invention utilizes a specific primer set to obtain the bovine ATP5PO gene fragment associated with normal and thawed bovine sperm motility, and uses specific SNP sites within this fragment as molecular markers, providing a theoretical basis and practical application for marker-assisted selection in cattle.

The application relates to the technical field of animal gene detection, and discloses an application method of using an IL1RAP gene as a molecular marker of a superovulation trait of a cow, which comprises the following steps: extracting a genomic DNA template from blood of a to-be-detected cow individual; using a forward primer and a reverse primer to combine the template to perform PCR amplification, obtaining an amplified fragment to obtain a PCR product; performing sequence determination on the PCR product, detecting single nucleotide polymorphism caused by a C to T base mutation existing at the 198th position of a DNA sequence to obtain a target genotype; and performing correlation analysis on the target genotype and the superovulation trait of the cow, and selecting the to-be-detected cow individual with the TT type when the target genotype is the TT type. The application realizes the application of the IL1RAP gene as the molecular marker by detecting single nucleotide polymorphism of the amplified fragment, performing correlation analysis on the target genotype and the ovulation trait, and screening the TT type individual, and the influence of the polymorphism on superovulation is determined.

The invention belongs to the technical field of animal gene engineering, genetic modification and biological medicine, and particularly relates to an HSD17B13 gene modified non-human animal, a construction method thereof and application of the HSD17B13 gene modified non-human animal in the technical field of biological medicine. The construction method comprises the step of replacing a nucleotide sequence of endogenous HSD17B13 of a non-human animal with a nucleotide sequence containing human HSD17B13 at an endogenous HSD17B13 locus of the non-human animal, so that the obtained human or chimeric (e.g., humanized) HSD17B13 protein expressed by the non-human animal can be used as an animal model for researching the functions of the HSD17B13 gene and screening disease treatment drugs related to the HSD17B13, and the human or chimeric (e.g., humanized) HSD17B13 protein expressed by the non-human animal can be used as an animal model for researching the functions of the HSD17B13 gene and screening the disease treatment drugs related to the HSD17B13. The important application value is realized.

The invention discloses a construction method and application of an IL22 gene humanized animal, and relates to the technical field of biology. The construction method is characterized in that a whole genome sequence of exons 1-5 of a non-human animal Il22 gene is replaced by a whole genome sequence of exons 1-5 of a human IL22 gene, and a non-human animal genome contains a human sequence shown as SEQ ID NO.4. The construction method comprises the following steps: firstly, constructing a human-derived sequence; an ideal in-vivo research platform is provided for deeply researching a human IL22 signal channel mechanism and screening tumor and immune disease treatment medicines.

This invention provides a method for site-specific replacement of the creatine kinase promoter in the Huaxi bovine gene promoter, belonging to the field of animal gene editing and genetic breeding technology. It also provides a method for enhancing the function of bovine muscle cells. FTO Gene expression reagents include crRNA for upstream site knockout, crRNA for downstream site knockout, a Cas12i nuclease expression vector, and an FTO homologous substitution vector; the FTO homologous substitution vector includes... FTO upstream homologous arm of gene promoter, MCK gene promoter and FTO Downstream homologous arm of the gene promoter. The reagent was used in West China bovine cells. FTO Application in gene promoter replacement. This invention can specifically improve the expression efficiency of genes related to fat development in muscle tissue, providing an efficient and precise technical means for the genetic improvement of the performance of beef from Western China.

The present application relates to the technical field of biological chemical industry, and particularly relates to an acid-responsive gene carrier, a construction method and application thereof.The present application provides acid-responsive supramolecular nanoparticles SNs based on star cationic polymer beta-CD-PDMAEMA and Pasp-benzoic imine-Ad host-guest interaction of adamantane modified polyaspartamide, and experimental results show that the mixing ratio (Ad / CD) of Pasp-benzoic imine-Ad and beta-CD-PDMAEMA, and the mixing ratio (N / P) of SNs and DNA can affect cell toxicity, transfection efficiency and phagocytosis efficiency, wherein the complex has high cell transfection endocytosis efficiency and stability when the N / P ratio is 20 and the Ad / CD is 2 or 4; based on this, the carrier can be used for expression, gene silencing and / or intracellular endocytosis of tobacco or other plant and animal genes, or for breeding improvement of tobacco and other plant and animal cells, or for gene therapy, and has a wide application prospect.

The invention belongs to the field of animal gene engineering, and relates to a Spy transposon system and a gene transfer method mediated by the Spy transposon system, the Spy transposon system comprises cgSpy and cvSpy transposon and transposase; through cell verification, the cgSpy and cvSpy vector systems can effectively mediate gene transfer, and have great application potential in transgenic animal preparation and gene therapy.

The invention belongs to the technical field of animal genetic engineering, and particularly relates to a method for increasing the growth speed of pigs. The method comprises the following steps that under the condition that an exogenous gene is not introduced, a coding region of a GH1 gene and a strong promoter of another endogenous high-expression gene are fused in vivo, the GH1 high-expression gene is generated, and the strong promoter is a PSMC5 gene promoter, an ACTA1 gene promoter, an ACTB gene promoter, an NME2 gene promoter or an RPL23 gene promoter. According to the method, pig varieties which grow rapidly can be created.

The invention provides a tent5d gene mutation and application thereof, and belongs to the technical field of molecular biology and medicine. The CDS sequence of the mutant gene of the tent5d gene is as shown in SEQ ID NO.1, and compared with the CDS sequence of a normal human TENT5D gene, the CDS sequence of the mutant gene of the tent5d gene has c.C555G (p.D185E) mutation. A male sterility animal model can be constructed by enabling corresponding sites of animal tent5d genes to be subjected to same mutation, when the animal model is a mouse model, the tent5d genes of a mouse are subjected to c.T555G (p.D185E) mutation to obtain a male sterility mouse model, and the mouse model can be used for accurately simulating pathogenic mutation of human genetic diseases. And a key experimental platform is provided for development of gene therapy, targeted drug screening and germ cell editing research. The invention has important significance in diagnosis and treatment of male infertility.

The invention relates to a construction method and application of an IgA nephropathy animal model, and belongs to the technical field of biology. The construction method comprises the step of injecting an effective dose of antisense gamma PNA capable of targeting an animal C1GALT1 gene into an experimental animal through veins. The method is easy and convenient to operate and high in modeling success rate, and a research model is provided for exploring IgA nephropathy occurrence and related mechanisms.