12 results about "Animal gene" patented technology

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.

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 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.

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.

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.

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 application belongs to the field of animal genetic engineering and molecular biotechnology, and particularly relates to FDXR The application of the expression activator of the gene in the preparation of a product for resisting porcine epidemic diarrhea virus infection. FDXR The nucleotide sequence of the gene is shown as SEQ ID NO. 1. The research result of the application shows that the expression level of the gene will affect the replication level of PEDV. FDXR When the expression of the gene in a host cell is inhibited, the replication of PEDV can be significantly promoted and the titer of the virus is increased. FDXR When the FDXR is overexpressed in the host cell, the replication of PEDV can be significantly inhibited and the titer of the virus is decreased. FDXR The experimental result not only shows that the gene can be used as a new target for the development of an antiviral drug for PEDV, but also provides a tool and material for the research on the molecular mechanism of the gene in the regulation of the replication of a pathogenic microorganism in a cell. FDXR The application belongs to the field of animal genetic engineering and molecular biotechnology, and particularly relates to

The application belongs to the technical field of animal genetic engineering, and provides a FETUB gene molecular marker related to the sperm vigor trait of a cow and application thereof, wherein the FETUB gene molecular marker is an SNP site on a FETUB gene fragment of the cow as shown in the sequence table SEQ ID NO:1, specifically, the 315th site of the FETUB gene fragment, the site has an A-G base mutation, resulting in the generation of a single nucleotide polymorphism of the gene. Based on the FETUB gene molecular marker, the genotype is determined by sequence determination, the genotype is associated with the normal and post-thawing vigor traits of the sperm of the cow, and the normal and post-thawing vigor of the sperm of individuals with different genotypes has a significant difference. The FETUB gene molecular marker provided by the application provides a theoretical basis and specific application for marker-assisted selection of the cow.

This invention discloses the application of the USP28 gene in the proliferation of porcine muscle stem cells, belonging to the field of animal genetic engineering. Resequencing data shows that this gene is selected in pig breeds with high meat production traits. Functional verification indicates that knocking down this gene in primary porcine muscle stem cells significantly reduces the mRNA expression level of the proliferation marker gene and significantly decreases the number of EdU-labeled proliferative positive cells. This invention clarifies the proliferation-promoting function of the USP28 gene, which can be used as a molecular marker or target gene for pork quality improvement and new breed breeding, providing an effective technical means to solve the problems of insufficient pork yield and genetic improvement of quality traits.

The application discloses a CRHR1 gene molecular marker for detecting a superovulation trait of a cow and an application method, belongs to the technical field of animal gene engineering, and has the nucleotide sequence length of 666bp of the obtained CRHR1 gene fragment, wherein a c-t base mutation exists at the 455th bp of the segment; the mutation site is used as a molecular marker, genotypes are determined through sequence determination, and the genotypes can be associated with the excellent superovulation trait of the cow. Analysis results show that the superovulation performance of individuals with different genotypes has significant differences. The application uses specific primers to obtain a partial CRHR1 gene segment related to the excellent superovulation trait of the cow, and uses a specific SNP site in the segment as a molecular marker, so as to provide a theoretical basis and specific application for marker-assisted selection of the cow.

The application belongs to the technical field of animal genetic engineering, and provides a DHX36 gene molecular marker related to the sperm vigor trait of a cow and application thereof, wherein the DHX36 gene molecular marker is a C-G base mutation at the 231st site of a bovine DHX36 gene fragment shown in the sequence table SEQ ID NO:1. Based on the DHX36 gene molecular marker, the genotype is determined by sequence determination, and the genotype is associated with the normal and post-thawing sperm vigor trait of the cow. The analysis result shows that the normal and post-thawing sperm vigor of individuals with different genotypes is significantly different. The application uses specific primers to obtain a bovine DHX36 gene fragment related to the normal and post-thawing sperm vigor trait of the cow, and uses a specific SNP site in the fragment as a molecular marker, thereby providing a theoretical basis and specific application for marker-assisted selection of the cow.