24 results about "Bovine muscle" patented technology

The invention discloses an enhancer Enh483 related to bovine muscle development and application of the enhancer Enh483 in promoting bovine myoblast proliferation and / or differentiation. According to the method, existing ATAC-Seq data and Chip-seq data in the proliferation stage and differentiation stage of buffalo myoblasts are utilized to visually analyze chromatin open areas through IGV to predict annotation enhancers, candidate enhancers Enh483 related to muscle development are screened out, and the enhancers have enhancer activity detected by a dual-luciferase vector report system; after the activity of Enh483 is inhibited, the proliferation and differentiation of buffalo myoblasts can be inhibited, which indicates that Enh483 can promote the proliferation and / or differentiation of buffalo myoblasts. According to the invention, an important theoretical basis is provided for an action mechanism of transcription regulation elements such as enhancers in muscle development, and data support is provided for buffalo breeding and strain breeding.

The invention relates to the technical field of gene engineering, in particular to application of a knock-down CDYL gene in improvement of cattle meat production performance. The invention provides an application of knocking down a CDYL gene in improving the differentiation capacity of cattle muscle satellite cells and / or improving the meat production performance of cattle. The gene ID of the CDYL gene is 539013. The 404th basic group A on the upstream of the CDYL gene transcription start site is deleted through a gene editing method, so that the CDYL knock-down gene editing cattle is created, compared with wild cattle and gene editing cattle, the myotube forming ability of muscle satellite cells is obviously enhanced, the number of myotube marker intermuscular line protein positive cells is obviously increased, the number of multinuclear myotubes is obviously increased, and the CDYL knock-down gene editing cattle has the advantages that the CDYL knock-down gene editing cattle can be used for preparing the CDYL knock-down gene editing cattle. And the weight and body size data (body height, cross height, body oblique length, chest circumference and abdominal circumference) of the gene edited cattle are greater than those of the wild cattle. According to the invention, high-yield breeding genes of beef cattle are enriched, and technical support is provided for cultivation of novel gene editing animals.

This application relates to the technical field of Yanhuang cattle breeding production, specifically to a molecular marker associated with the muscle fatty acid content of Yanhuang cattle and its application. This molecular marker includes a nucleotide sequence formed by a C>T mutation at position 20,849,921 bp on chromosome 20 of the bovine reference genome GCF_002263795.3. This molecular marker is closely associated with the muscle fatty acids of Yanhuang cattle and can be used in the breeding of high-quality Yanhuang cattle rich in high-quality fatty acids.

According to the application of the HTATSF1 gene in regulation and control of cattle muscle growth and development and breeding, the influence of the HTATSF1 gene on muscle growth and development and the correlation between HTATSF1-CNV and cattle growth traits are verified, a reliable molecular marker is provided for cattle seed selection and breeding, and help is provided for cattle variety improvement. A foundation is laid for association between copy number variation and growth traits of the cattle HTATSF1 gene, cattle molecular marker-assisted selection breeding work can be accelerated, the method is simple, rapid and convenient to popularize and apply, and a complete solution from fundamental research to industrial application is provided for improvement of cattle germplasm resources.

The invention discloses an edible beefsteak scaffold based on vegetable protein and a construction method thereof, and belongs to the fields of tissue engineering, material science, food science and biology. The edible beefsteak scaffold is a 3D printing scaffold based on vegetable protein, combined with gellan gum and sodium alginate, and is a cell culture meat bionic beefsteak composed of a bovine fibroblast and bovine muscle satellite cell co-culture system dynamically constructed by using a rotary culture system. The selected materials are all edible protein and polysaccharide, have high biocompatibility and safety, and are wide in source; the mode of dynamically constructing the bionic beefsteak can effectively reduce the formation of local necrotic areas, and the bionic beefsteak has excellent physical characteristics, has texture structures and flavor characteristics similar to those of real beefsteak, is sterile in surface, and can significantly reduce food sanitation and safety problems. The scheme provided by the invention has remarkable advantages in the aspects of food safety, texture regulation and control, sensory simulation and the like, and provides important technical support for the field of cell culture meat.

The present invention relates to the field of genetic engineering technology, and particularly to the application of knocking down CDYL genes in improving the meat production performance of cattle. The present invention provides the application of knocking down CDYL genes in improving the differentiation ability of bovine muscle satellite cells and / or improving the meat production performance of cattle, and the gene ID of the CDYL gene is 539013. The present invention deletes the 404th base A upstream of the transcription start site of the CDYL gene by gene editing, thereby creating a CDYL gene-edited cattle with knocked-down gene. Compared with wild-type cattle, the myotube formation ability of muscle satellite cells in gene-edited cattle is significantly enhanced, the number of desmin-positive cells, a myotube marker, is significantly increased, the number of multinucleated myotubes is significantly increased, and the body weight and body size data (body height, rump height, body slant length, chest girth and abdominal girth) of gene-edited cattle are all greater than those of wild-type cattle. The present invention enriches the breeding genes for high-yield beef cattle and provides technical support for the cultivation of new gene-edited animals.

The invention discloses a muscle cell efficient growth gene knock-in cell line construction method based on a CRISPR-Cas9 system, and belongs to the crossing field of gene editing and biotechnology. Through a CRISPR-Cas9 mediated gene targeting insertion technology, fixed-point integration of muscle efficient growth related functional genes is realized in bovine fetal fibroblasts, and a gene overexpression cell line with stable hereditary characteristics is successfully constructed. The cell line can be used as a nuclear donor, and precise creation of high-muscle-production-performance gene editing beef cattle is realized by combining a somatic cell nuclear transfer technology and an embryo transfer technology. The key technical support is provided for beef cattle molecular design breeding by directionally enhancing the cattle muscle development core regulation and control network. The genetic improvement new material cultivated based on the technology is expected to significantly improve the beef cattle muscle growth efficiency and industrial economic benefits, and shows important application value and industrialization prospects in the field of agricultural animal genetic engineering.

This invention relates to the fields of stem cell culture and serum-free culture media, specifically to a serum-free culture medium for the proliferation of bovine muscle stem cells and its applications. The serum-free culture medium is based on a serum-free basal culture medium, to which food-derived small molecule compounds are added. Through further combination optimization, four food-derived small molecule compounds with synergistic effects have been obtained. This serum-free culture medium can significantly promote the proliferation of bovine muscle stem cells, effectively maintain cell proliferation activity, and cells passaged using this serum-free culture medium can still maintain good differentiation potential. This provides a stable, sufficient, and safe cell source for the industrial production of cultured meat, which is beneficial for the large-scale production of bovine muscle stem cells.

The present invention relates to a molecular marker related to the level of betaine content in cattle muscle, located at the 102029509th base from the 5′ end on chromosome 9 of the cattle reference genome ARS-UCD1.2. The present invention describes a primer pair for detecting the aforementioned molecular marker, a kit, and its application in identifying the level of betaine content in cattle muscle, screening cattle breeds, or cattle genetic breeding. The relative abundance of betaine in cattle muscle of individuals with AA genotype is 0.90, and that of individuals with GG genotype is 1.45. By selecting AA-type individuals, cattle with higher muscle betaine content can be obtained. The present invention effectively solves the problem that the betaine content in cattle muscle cannot be measured in vivo in actual production, reduces breeding costs, and effectively increases or decreases the betaine content in cattle muscle in actual production. The method has high accuracy, low detection cost, and can realize automated detection, and has high practical application value in cattle breeding.

The invention discloses a cell co-culture device, which relates to the field of cell culture. The device is composed of a dish body and a dish cover. The dish body is a hollow structure with an upper opening. The dish cover is provided at the opening of the dish body. The dish body is provided with an isolation ring. The isolation ring divides the interior of the dish body into region I and region II. The isolation ring contains a crawling sheet. The device can be used to realize direct contact co-culture of bovine muscle cells and fat cells, and single cells (muscle cells or fat cells) on the crawling sheet can be collected for subsequent transcriptome and proteome analysis and processing to study the effects on muscle cells or fat cells in a co-culture environment. Therefore, it is beneficial to further explore the interaction between muscle cells and fat cells. It has been confirmed through multiple observations that the muscle cells and fat cells cultured in this way have high survival efficiency, good vitality, obvious cell characteristics, and are consistent with the morphology of muscle cells and fat cells cultured alone. It is a good in vitro research model with certain promotion and research value.

The invention discloses a method, a device, equipment and a medium for screening and detecting key genes of beef produced by Xianan cattle. The method comprises the following steps: preprocessing genome data, apparent group data and beef phenotype data; screening specific mononucleotide polymorphic sites differentiated from other beef cattle varieties through a dynamic population differentiation index threshold value, performing function annotation, performing correlation analysis on the sites subjected to the function annotation and meat production phenotype data through a three-order analysis model combined with captured data verification to obtain candidate key genes, and then identifying the candidate key genes according to the candidate key genes. Detecting the relative expression quantity of the candidate key gene in the muscular tissue of the Xianan cattle through quantitative polymerase chain reaction, and analyzing and verifying the relevance between the candidate key gene and the meat production phenotype in combination with Pearson correlation; specific polymerase chain reaction primers are designed for the key genes passing verification, and detection of the key genes is achieved through fluorescent quantitative polymerase chain reaction.

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 invention provides a beef cattle MSTN gene promoter function optimization design method based on deep learning and model adaptation. The beef cattle MSTN gene promoter function optimization design method comprises the following steps: taking RNA-seq expression quantity of an MSTN gene as a supervision label, and taking high-dimensional epigenetic signal prediction data obtained by inputting a promoter region sequence into a pre-trained Enformer deep learning model as an input feature; training a machine learning adapter model; when the prediction precision of the adapter model reaches a preset threshold value, determining the adapter model as a verification model; constructing a promoter variant sequence library of the MSTN gene by adopting a multi-strategy variant generation method; the promoter activity is verified by a reporter gene expression verification method aiming at the promoter sequence which ranks in the top in the variant sequence library. The invention also discloses an enhanced promoter with a sequence as shown in SEQ ID NO.4 and a weakened promoter with a sequence as shown in SEQ ID NO.2 or 3.

Provided is a technique with which it is possible to improve the proliferation rate of cells in culture in which microcarriers are used. A cell line is established by introducing, into bovine progenitor cells, the following four genes: CDK4 (R24C), TERT, CCND1, and p53 that lacks a DNA binding domain.

The application belongs to the field of cell culture, and discloses a bovine muscle stem cell in-vitro separation and culture method, a culture medium and application thereof. The muscle stem cell in-vitro culture method of the application is to culture muscle stem cells by using a cell culture medium of non-animal source hydrolysate. The application further discloses a culture medium for culturing the muscle stem cells. By using the culture medium of the application, muscle stem cells with ideal cell quantity and cell morphology can be obtained on the basis of no or less serum addition, and the muscle stem cells can still maintain stemness and differentiation potential after continuous passage for multiple times. The application utilizes the in-vitro cultured muscle stem cells for the field of cell culture meat, and has a wide application prospect.

This invention provides a technology that can improve the proliferation rate of cells in cultures using microcarriers. [Solution] The cell line is established by introducing the following four genes: CDK4(R24C), TERT, CCND1, and p53 lacking the DNA binding domain into bovine progenitor cells.

The invention belongs to the technical field of biology and cells, and discloses application of a circAAGAB gene in regulation and control of proliferation and differentiation of bovine skeletal muscle satellite cells, the circAAGAB gene truly exists and is in a ring shape, and the nucleotide sequence of the circAAGAB gene is shown as SEQ ID NO.1. The circAAGAB gene has a nucleotide sequence shown as SEQ ID NO.2. The circAAGAB gene has a nucleotide sequence shown as SEQ ID NO.1. According to the method disclosed by the invention, the proliferation and myogenic differentiation process of the bovine muscle satellite cells can be effectively regulated and controlled by regulating the expression of circAAGAB. A new enlightenment is provided for researching muscle development and differentiation, and an important reference is provided for exploring a muscle growth and development mechanism and improving the yield and quality of beef. In addition, a good foundation is laid for deeply discussing the action mechanism of circAAGAB in the muscle development process, and theoretical support is provided for rapid breeding and meat quality improvement of new varieties of high-quality beef cattle.

The invention relates to the field of stem cell culture and serum-free culture media, in particular to a serum-free culture medium for bovine muscle stem cell proliferation and application of the serum-free culture medium. According to the serum-free culture medium, food-borne small molecule compounds are added on the basis of a serum-free basic culture medium, and four food-borne small molecule compounds with a synergistic effect are obtained through further combinatorial optimization. The serum-free culture medium can significantly promote proliferation of bovine muscle stem cells and effectively maintain the activity of cell proliferation, cells subcultured by the serum-free culture medium can still maintain good differentiation potential, a stable, sufficient and safe cell source is provided for industrial preparation of cell culture meat, and the serum-free culture medium has good application prospects. The large-scale production of the bovine muscle stem cells is facilitated.

This invention relates to the field of bioengineering technology, specifically to the application of the AIFM2 gene in regulating intramuscular fat deposition in bovine muscle. This invention discloses the function and application of the apoptosis-inducing factor mitochondrial-associated 2 (AIFM2) gene in regulating intramuscular fat deposition in bovine muscle. Cellular experiments and animal model experiments have confirmed that AIFM2 gene overexpression significantly promotes adipogenic differentiation of CD3 / CD4 / PDGFR fibroblast / adipogenic progenitors (FAPs), increases lipid droplet accumulation, and promotes lipid deposition in muscle tissue and increases serum triglyceride levels in mice. Its mechanism of action is through the activation of the expression of key adipogenic differentiation genes such as PPARγ, C / EBPα, ADIPOQ, and FABP4 at the transcriptional and protein levels, thereby regulating lipid metabolism and regional deposition. This invention confirms that the AIFM2 gene can promote intramuscular fat deposition in bovine muscle, providing a reference for bovine molecular breeding.

The invention discloses antisense l ncRNA CFL1-AS1 related to bovine muscle growth and development and application thereof, and belongs to the technical field of molecular biology, through high-throughput sequencing, differentially expressed l ncRNA TCONS00184439 is screened from adult bovine muscle tissue, RACE cloning is performed to obtain a full-length sequence 616bp, genomic sequences are compared, the positions of the genomic sequences and the antisense strand of a CFL1 gene are overlapped on a genome, and the antisense 1 ncRNA CFL1-AS1 related to bovine muscle growth and development is obtained. The antisense lncRNA is named as CFL1-AS1 according to the naming of the long-chain non-coding RNA; the CFL1-AS1 has specific low expression in adult cattle muscle tissues, the CFL1-AS1 is up-regulated to promote myoblast proliferation, myoblast differentiation is dynamically regulated and controlled, and the CFL1-AS1 participates in the muscle growth and development process; the CFL1-AS1 overcomes the defects of the antisense l ncRNA in muscle growth and development, and provides a new molecular marker and a new breeding idea for genetic improvement and efficient breeding of beef cattle.

The invention discloses an immortalized bovine muscle-derived stem cell line as well as a construction method and application thereof, and belongs to the technical field of stem cells and biological cell lines. The bovine muscle-derived stem cell line is characterized in that telomerase reverse transcriptase (TERT) and cyclin dependent kinase (CDK) genes are overexpressed in a mode of lentivirus packaging and infection, a cell strain which is single in source and stably expresses a target gene is obtained through monoclonal screening, gene verification and functional verification, and the verification shows that the cell line overexpresses the CDK4 and TERT genes, so that the cell line can be used for preparing the bovine muscle-derived stem cell line with the stable expression of the target gene. The monoclonal cell strain can stably proliferate and maintain good differentiation characteristics. The immortalized bovine muscle-derived stem cell line is prepared, and a good seed cell resource is reserved for commercialized production of cultured meat.

The invention belongs to the technical field of biology, cell and tissue engineering, and discloses application of an FGF16 gene in regulation and control of bovine skeletal muscle satellite cell differentiation, and the nucleotide sequence of the FGF16 gene is SEQ ID NO.1. According to the method disclosed by the invention, the proliferation and myogenic differentiation process of the bovine muscle satellite cells can be effectively regulated and controlled by regulating the expression of the FGF16. A new enlightenment is provided for researching muscle development and differentiation, and an important reference is provided for exploring a muscle growth and development mechanism and improving the yield and quality of beef. In addition, a good foundation is laid for deeply discussing the action mechanism of the key FGF16 protein in the muscle development process, and theoretical support is provided for rapid breeding and meat quality improvement of new varieties of high-quality beef cattle.

The invention provides a method for site-specific replacement of a West China cattle gene promoter with a creatine kinase promoter, and belongs to the technical field of animal gene editing and genetic breeding. The invention discloses a reagent for improving FTO gene expression in bovine muscle cells. The reagent comprises crRNA for upstream site knockout, crRNA for downstream site knockout, a Cas12i nuclease expression vector and an FTO homologous replacement vector, the FTO homologous replacement vector comprises an FTO gene promoter upstream homologous arm, an MCK gene promoter and an FTO gene promoter downstream homologous arm. The reagent is applied to FTO gene promoter replacement in western China cattle cells. According to the method, the expression efficiency of fat development related genes in muscle tissues can be specifically improved, and an efficient and accurate technical means is provided for genetic improvement of the performance of the beef in the western China.

The application discloses a bovine muscle development related enhancer Enh483 and application thereof in promoting bovine myoblast proliferation and / or differentiation. The application predicts and annotates the enhancer by IGV visual analysis of chromatin open regions by using existing buffalo myoblast proliferation period and differentiation period ATAC-Seq data and Chip-seq data, screens out a candidate enhancer Enh483 related to muscle development, and detects the enhancer activity by a dual luciferase vector report system. After the Enh483 activity is inhibited, the buffalo myoblast proliferation and differentiation are inhibited, which indicates that the Enh483 can promote the buffalo myoblast proliferation and / or differentiation. The application provides an important theoretical basis for the mechanism of the transcriptional regulatory elements such as enhancers in muscle development, and provides data support for buffalo breeding and strain selection.