Product
Patsnap Eureka
For R&D, Patsnap Eureka makes reading and utilizing patents & technical documents easy.
Patsnap Eureka AIR
Designed for self-driven R&D workflows. Generate viable solutions, solve complex R&D challenges, empower your innovation with AI.
Patsnap Eureka Materials
Designed for material experts only. Revolutionize your material R&D, from search, analyze, to developing new materials.
Feature
TechResearch
Generate reliable direction feasibility study reports for your R&D in just a few steps.
TechSeek
Discover and master advanced knowledge NOW. Basics, ideas, possibilities, all at once.
TechMind
As an expert in R&D Theories, TechMind can generates customized viable solutions instantly.
TechRisk
Analyze your overall solution with one click, know your potential R&D risks in advance.
TechMonitor
Get weekly tech updates, stay abreast of the latest tech innovations and key insights.
Patents
Literature
Hiro is an intelligent assistant for R&D personnel, combined with Patent DNA, to facilitate innovative research.
42 results about "Somatic Cell Nuclear Transfer Technique" patented technology
Efficacy Topic
Property
Owner
Technical Advancement
Application Domain
Technology Topic
Technology Field Word
Patent Country/Region
Patent Type
Patent Status
Application Year
Inventor
Somatic cell nuclear transfer (SCNT), technique in which the nucleus of a somatic (body) cell is transferred to the cytoplasm of an enucleated egg (an egg that has had its own nucleus removed).
Method of cloning reproductive and respiratory syndrome resisting pig
Owner:CHINA AGRI UNIV
Overexpression porcine co-stimulatory 4-1BB vector and application thereof
InactiveCN105087620AHigh copy numberLower activation thresholdVector-based foreign material introductionAnimal husbandryInteinEmbryo
The invention provides an overexpression porcine co-stimulatory 4-1BB vector and application thereof. PCR (polymerase chain reaction) amplification is performed on a left homologous arm and a right homologous arm of an intron 1 of a rosa26 gene, a 4-1BB regulatory sequence and an OCT4 specific promoter; the left homologous arm, a 4-1BB expression cassette, LoxP locus-contained Cre and Neo expression cassettes, the right homologous arm and negative selection DTA diphtheria toxin are connected in sequence to obtain a 4-1BB homologous recombinant vector p4BOCNDR; the vector and a CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR-associated) targeting vector of sgRNA (small guide ribonucleic acid) containing the intron 1 of the specific targeting porcine rosa26 gene are transferred together into a porcine fetus fibroblast; by taking a positive cell as a donor cell and an oocyte as a recipient cell, a cloned embryo is obtained through a somatic cell nuclear transfer technique; the cloned embryo is transplanted into a porcine uterus for fetation to obtain a transgenic pig integrating a 4-1BB gene at the fixed point of a first intron of the rosa26 gene and automatically deleting a marker gene.
Owner:CHINA AGRI UNIV
Method for preparing MC4R gene knockout pig
The invention relates to the field of genetic engineering and genetic modification, in particular to a method for obtaining an MC4R gene knockout pig by using a CRISPR / Cas9 system for editing MC4R genes through a somatic cell nuclear transfer technology. sgRNA is designed by aiming at two sites (a 6133-6152bp sequence and a 7127-7146 bp sequence in a CDS region of the MC4R gene) in the CDS region of the pig MC4R gene for the first time; in addition, the CRISPR / Cas9 system is used for simultaneously cutting the two sites; the large segment deletion of the MC4R gene is realized; in addition, the gene knockout individual pig subjected to large segment deletion is obtained; a feasible method is provided for studying the pig MC4R gene.
Owner:CHINA AGRI UNIV
Method for preparing high muscle content and hypertrophic cardiomyopathy model cloned pig
InactiveCN105463027AImprove convenienceShorten the timeNucleic acid vectorFermentationHypertrophic cardiomyopathyNuclear transfer
The invention provides application of a Trim63 gene in preparation of a high muscle content and hypertrophic cardiomyopathy model cloned pig. Somatic cells of the Trim63 gene of a modified pig are utilized as nuclear transfer donor cells, oocytes are utilized as nuclear transfer recipient cells, and a cloned embryo is obtained through the somatic cell nuclear transfer technology. The cloned embryo is transferred into a pig uterus to gestate to obtain the Trim63 gene modified high muscle content and hypertrophic cardiomyopathy model cloned pig, artificial intervention methods, such as operation and so on for the cloned pig are not needed any more, and the efficiency of establishment of a disease model is improved. According to the high muscle content and hypertrophic cardiomyopathy model cloned pig, paired Cas9n targeting vectors are utilized for the first time to perform gene editing for large animals. The method is low in cost, sharply shortens the time for obtaining a homozygote pig and lays a foundation for gene function research and the disease model establishment for the large animals by utilizing the CRISPR / Cas9 technology.
Owner:CHINA AGRI UNIV
Production method for Fbxo40 gene knockout pigs
InactiveCN105821049AConvenient researchLow costNucleic acid vectorFermentationDiseaseAgricultural science
The invention provides a production method for Fbxo40 gene knockout pigs. The production method comprises the steps that a CRISPR-Cas9 targeting vector and a PGK-Neo resistant gene are jointly transfected into a porcine embryonic fibroblast to obtain a G418-resistant positive monoclonal cell, an Fbxo40 gene in the positive monoclonal cell generates insertion or deletion mutation, a reading frame generates frame shift and stops in advance, then the obtained cell is cloned to serve as a donor cell for nuclear transfer, and an oocyte serves as a receptor oocyte for nuclear transfer; cloned embryos are obtained through a somatic cell nuclear transfer technique, the high-quality cloned embryos are transferred into the fallopian tube of an oestrous sow, and the Fbxo40 gene knockout pigs are obtained through whole-period development. According to the production method, the Fbxo40 knockout pigs are efficiently obtained through a CRISPR-Cas9 gene editing technique at the low cost, and animal models are supplied to research of muscle development and diseases related to the muscles.
Owner:CHINA AGRI UNIV
CRISPR/Cas9 technology mediated down producing goat VEGF gene fixed-point knock-in method
ActiveCN108570479AGrowth and development effectsAvoid combiningHydrolasesStable introduction of DNANuclear transferHIV receptor
The invention provides a method for finishing down producing goat VEGF gene fixed-point knock-in mediated by a CRISPR / Cas9 system. The method comprises the following steps: establishing a gRNA expression vector and a VEGF homologous recombination vector on the basis of the CRISPR / Cas9 system according to a CCR5 gene sequence of down producing goats; then jointly transferring three optimized vectors into fibroblasts of down producing goat fetuses to obtain VEGF gene fixed-point knocked-in positive cells; and preparing VEGF gene fixed-point integrated down producing goats by using a somatic nuclear transfer technology. The established targeting vector based on the CRISPR / Cas9 system provides a simple, rapid and safe path for fixed-point knock-in of goat VEGF genes. The method does not involve any screening marker genes in a cell line screening process, therefore, the safety of transgenic animals is greatly improved, and the method has high value on research of genetic breeding and gene functions of down producing goats.
Owner:INNER MONGOLIA UNIVERSITY
Reconstructed ovum of albinism model pig and construction method thereof, and construction method of model pig
ActiveCN104263754AVector-based foreign material introductionAnimal husbandryHypersensitive responseAlbinism
The invention relates to a reconstructed ovum of an albinism model pig and a construction method thereof, and a construction method of the model pig. According to the construction method of the reconstructed ovum, tyrosinase is knocked out to obtain the reconstructed ovum of the albinism model pig. The combination of a CRISPP / Cas9 gene knock-out technique and a somatic nucleus transplantation technique proves that the method for constructing the gene modified pig has high feasibility. The obtained TYR gene knock-out pig has the typical characteristics of albinism, can provide a reliable animal model for researching human albinism, can be applied to toxicity and anaphylaxis detection and many other aspects, and is hopeful to become a standardized experimental animal for albinism-like mice.
Owner:广州中科飞豚生物科技有限责任公司
Preparation method of liver cells with low expression or no expression of PERV
InactiveCN105154471AAchieve knockoutReduced activityVector-based foreign material introductionForeign genetic material cellsNuclear geneSomatic cell
The invention discloses a preparation method of liver cells with low expression or no expression of PERV. The preparation method includes the following steps: (1), respectively preparing a recombinant vector containing siRNA applied to RNA interference technology, a recombinant vector containing gRNA applied to CRIPRS / Cas9 technology and a vector containing Cas9 applied to CRIPRS / Cas9 technology; (2), jointly transfecting the three vectors to porcine somatic cells, culturing the transfected porcine somatic cells, and screening and identifying to obtain monoclonal genetically-modified porcine somatic cells; (3), utilizing somatic cell nuclear transfer technology to obtain genetically-modified pig containing nuclear genes of the genetically-modified porcine somatic cells; (4), extracting liver cells of the genetically-modified pig. According to the preparation method, knockout before transcription and inhibition after transcription of PERV genes are realized, and the technical scheme is effective and feasible.
Owner:ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
Method for producing recombinant human BChE (butyrylcholinesterase) from transgenic animals by using gene knock-in and nuclear transfer technologies
The invention discloses a method for producing recombinant human BChE (butyrylcholinesterase) from transgenic non-human mammals by using gene knock-in and nuclear transfer technologies, and particularly provides a donor construct used for specific expression of a mammary gland. The construct integrates DNA (deoxyribose nucleic acid) coding sequences of human BChE to a specific expression protein gene locus of the mammary gland in targeted manner and performs successful transfection on non-human mammal embryo fibroblasts, juvenile and adult somatic cells or embryonic stem cells, clone is performed by using a somatic cell nuclear transfer technology so as to obtain a new individual, and the carried human BChE gene enables the mammary gland of a non-human mammal to express stably and secrete holoenzyme or a monomer of the human BChE. Recombinant protein produced with the method can be used for preventing and treating nerve poison and organic phosphorus compound poisoning as well as apnea caused by cocaine poisoning and succinylcholine, and for detecting and removing residues of organic phosphorus compounds on vegetables, other crops, various article layers and the soil.
Owner:SHANGHAI JENOMED BIOTECH CO LTD
CD28 gene overexpression vector and application thereof
ActiveCN105112449AHigh copy numberLower activation thresholdVector-based foreign material introductionAnimal husbandryGestationIntein
The invention provides a CD28 gene overexpression vector and application thereof. A target biological genome serves as a template, left and right homologous arms of a first intron of an ROSA26 gene, a CD28 gene expression frame and an OCT4 gene promoter are amplified through PCR (polymerase chain reaction), and the left homologous arm, the CD28 gene expression frame, LoxP-locus-containing Cre expression frame and Neo gene expression frame both regulated by the OCT4 gene promoter, the right homologous arm and a negative selection marker DTA are connected sequentially and are constructed to an eukaryotic expression vector to obtain the overexpression vector. The overexpression vector and a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas (CRISPR-associated) 9 target vector containing a first intron of a specific target pig ROSA26 gene are transplanted into a fibroblast of a pig fetus, a positive cell serves as a donor cell, an oocyte serves as a recipient cell, and a cloned embryo is obtained according to a somatic cell nuclear transfer technology; the cloned embryo is transplanted into the uterus of a pig for gestation to obtain a transgenic pig with a CD28 gene which is integrated at a fixed point in the first intron of the ROSA26 gene, wherein a marker gene of the transgenic pig is knocked out automatically.
Owner:CHINA AGRI UNIV
Transgene pig for preparing animal model for human diseases and culturing method thereof
InactiveCN101570763AVector-based foreign material introductionAnimal husbandrySomatic cellGenetic engineering
The invention provides a transgene pig for preparing an animal model for human diseases and a culturing method thereof. An inducible expression Cre recombinase expression vector pET28a-Mxl-Cre-BGHpolyA-FRT2neoR is constructed by a genetic engineering method, and can express Cre recombinase after the expression vector is induced by interferon inducer poly IC (poly I:C) so as to be used for knocking out a target gene; porcine mechanocyte capable of inducibly expressing the Cre recmbinase can be screened through technology of cell transfection and cell screening; and the pig is cloned by the technology of somatic cell nuclear transplantation. The cloned pig can be used for preparing the animal model for knocking out genes of human diseases.
Owner:JILIN UNIV
Method for obtaining lymphoma minipig disease model by knocking out P53 genes
The invention relates to a method for obtaining a lymphoma minipig disease model by knocking out P53 genes, and belongs to the field of medical animal disease models. The method mainly comprises the steps that TALEN target spots of the P53 genes are designed, P53 gene knockout TALEN plasmid pCAG-pP53-L and pCAG-pP53-R of the P53 genes are assembled, TALEN plasmid is transfected and screened to obtain a P53 gene knockout positive cell line, reconstructed embryos are constructed based on the somatic cell nuclear transplantation technology and transplanted into the body of a surrogate sow to continue to develop, and a piglet with the P53 genes knocked out is obtained; the function of the P53 genes is verified by detecting the expression level of RNA and protein of the P53 genes in tissue of the cloned piglet and downstream related genes, and lymphoma in the tissue of the piglet is identified through histopathology and an immunohistochemical method. The built lymphoma disease model has great significance in research and development of occurring, forming and medicine of human lymphoma, and the reliable model is provided for research and treatment of human lymphoma diseases.
Owner:魏红江 +1
Expression vector for piggyBac transposon, and transgenic pig and construction method thereof
InactiveCN103468732AHigh simulationEasy Vector ReconstructionVector-based foreign material introductionAnimal husbandryAgricultural scienceTransformation efficiency
The invention discloses an expression vector for a piggyBac transposon and a transgenic pig and a construction method thereof, belonging to the field of gene engineering. The expression vector is constructed by connecting a fusion fragment containing a loxP-Neo / EGFP gene expression cassette to pCyL50 plasmid. The vector is introduced into the genome of a pig, and somatic cell nuclear transfer technology is employed to produce the transgenic pig. The expression vector provided by the invention has the piggyback transposon, greatly improves transformation efficiency, changes a common plasmid series recombination transgene integration model into a single-site single-copy integration model and better simulates the internal environment of a biological gene. The construction method for the transgenic pig provides a novel approach for preparation of a variety of transgenic animals and a simple, convenient and highly efficient approach for fabrication of biological models of transgenic animals.
Owner:SOUTH CHINA AGRI UNIV
Expression vector resisting porcine circovirus type 2 (PCV2) and transgenic pig, and construction methods thereof
InactiveCN103468733AImprove efficiencyHigh simulationFermentationVector-based foreign material introductionTransformation efficiencyPorcine circovirus
The invention discloses an expression vector resisting PCV2 and a transgenic pig and construction methods thereof, belonging to the field of gene engineering. The expression vector is constructed by respectively connecting an H1 promoter and a pPB-sh2 sequence to a pUC57-H1 plasmid and then connecting a fusion fragment containing a loxP-Neo / EGFP gene expression cassette to pPB-H1-sh2. The vector is introduced into the genome of a pig, and somatic cell nuclear transfer technology is employed to produce the transgenic pig. The expression vector provided by the invention has a piggyback transposon, greatly improves transformation efficiency, changes a common plasmid series recombination transgene integration model into a single-site single-copy integration model and better simulates the internal environment of a biological gene. The constructed transgenic pig degrades PCV2mRNA through siRNA synthesized by itself, thereby fundamentally inhibiting PCV2 infection.
Owner:SOUTH CHINA AGRI UNIV
Targeting vector for pig GHR gene knock-out and applications thereof
The invention belongs to the technical field of biology, and specifically discloses a targeting vector for pig GHR gene knock-out and applications thereof. The targeting vector is a sgRNA expression vector based on a CRISPR / Cas system; wherein the target sequences of sgRNA are arranged on the 10th exon and 3'UTR of a pig GHR gene. The provided targeting vector is applied to targeting knock-out ofpig GHR gene, and the knock-out efficiency can reach 12.7%. After the GHR gene is successfully knocked out, somatic cells can be used to establish a GHR gene knock-out pig model by a somatic cell nuclear transplanting technology, and materials are provided for the research of physiological action and molecular mechanism of GHR gene.
Owner:CHINA AGRI UNIV
Reconstructed oocyte of deaf model pig, construction method of reconstructed oocyte, deaf model pig, and construction method and application of deaf model pig
PendingCN110468154AAvoid phenotypic differencesReliable Large Animal ModelsCompounds screening/testingHydrolasesDiseaseEmbryo transfer
The invention discloses a reconstructed oocyte of a deaf model pig and a construction method of the reconstructed oocyte, and also discloses the deaf model pig and a construction method and application of the deaf model pig. According to the reconstructed oocyte of the deaf model pig, the construction method of the reconstructed oocyte and the construction method of the deaf model pig, by knockingout a pig OSBPL2 gene, the reconstructed oocyte of a deaf mini-pig is obtained, and in combination with a CRISPR / Cas9 gene editing technology, a somatic cell nuclear transfer technology and an embryotransfer technology, high efficiency and feasibility of the construction method for the genetically engineered deaf model pig are proven. Through the construction method of the deaf model pig, genotypes and deafness phenotypes of human OSBPL2 gene mutations can be precisely duplicated, the pig of which an OSBPL2 gene is knocked out has typical characteristics of human autosomal dominant hereditary hearing loss (DFNA67), therefore, a reliable big animal model can be provided for disease research of the human hereditary hearing loss, and the deaf model pig can be applied to inner-ear gene repairing, inner-ear hair cell regeneration, hearing reconstruction and the like.
Owner:NANJING MEDICAL UNIV +1
Building method of donor pig suitable for xenotransplantation of tissue like islet cells and skin
PendingCN110373389AReduced responseReduce functionImmunoglobulin superfamilyGenetic engineeringAnimal biotechnologyIslet cells
The invention discloses a building method of a donor pig suitable for xenotransplantation of tissue like islet cells and skin and belongs to the technical field of animal biology. The building methodincludes: on the basis of porcine fetal fibroblast, utilizing CRISPR / Cas9 gene editing technology to knock out three genes GGTA1, CMAH and B4GalNT2 at the same time, and inserting three humanized modifying genes hCD55, hCD59 and hCD46 in a fixed point manner; combining somatic cell nuclear transfer technology to build an xenotransplantation donor base pig modified by the above six genes; on the basis of the base pig, knocking out gene B2M, and inserting three modifying genes hCD47, A20 and CTLA4-Ig in a fixed point manner at the same time; utilizing the somatic cell nuclear transfer technologyto build a islet cell and skin xenotransplantation donor pig modified by multiple genes; performing functional verifying on the donor pig to obtain the donor pig suitable for islet cell and skin xenotransplantation. A brand-new research concept and method are provided for developing the donor pig suitable for xenotransplantation of the islet cells and the skin, and clinical conversion and application of pigs serving in heterogeneous islet cell and skin transplantation are promoted.
Owner:YUNNAN AGRICULTURAL UNIVERSITY
Method and system for somatic cell nuclear transfer
InactiveUS20120142107A1Promote resultsIncrease the number of cellsGenetic engineeringFermentationNuclear transferEmbryo
Provided is a method for nuclear transfer. An exogenous donor nucleus is introduced into an enucleated oocyte and one or more enucleated sperm cells or one or more enucleate sperm cell fractions are introduced into the oocyte. The one or more sperm cells or one or more sperm cell fractions may be introduced into the oocyte either before, after, or simultaneously, with the donor nucleus. Also provided are cells and embryos produced by the method.
Owner:MAZOREL
Method for preparing and producing tissue type activator of plasminogen from mutant of transgene goat by nuclear transplantation
InactiveCN1659959ATo achieve the purpose of producing t-PAmImprove featuresFused cellsFermentationPhysiologyTissues types
A process for preparing transgenic she-goat to secrete human histotypic plasminogen activator mutant by nuclear transplant method features that the human histotypic plasminogen activator mutant regulated by the regulator sequence of beta-casein can be expressed in the milk gland of transgenic she-goat. It includes configuring carriers pGBCN-tPAm, pGBC2-tPAm and pGBC4-tPAm, transfecting the fibroblast of milk she-goat fetus, and cloning by nuclear transplanting.
Owner:INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
Method for breeding transgenic buffalos by using somatic cell nuclear transfer technology
InactiveCN102250954AFermentationVector-based foreign material introductionWater buffaloFull Term Pregnancy
The invention discloses a method for breeding transgenic buffalos by using somatic cell nuclear transfer technology, which comprises: obtaining buffalo fetal fibroblasts (BFF) by separation from buffalo fetuses and culture, implanting a transgenic vector (pCE-EGFP-IRES-neo), which carries a neomycin resistant gene and an enhanced green fluorescent protein (EGFP) gene, into the BFF by an electroporation process, and selecting BFF into which the EGFP gene is transferred by using G418; transplanting the BFF into which the EGFP gene is transferred into denucleated oocytes of the buffalo to construct cloned embryos, treating the cloned embryos for 5 minutes with ionomycin at a concentration of 5 mu mol / L, culturing cloned embryos in 6-dimethyl-aminopurine at a concentration of 2mmol / L for 3 hours, performing chemical activation treatment, co-culturing the cloned embryos in granulose cell monolayer liquid drops for 6 to 7 days, and obtaining the transgenic cloned blastaea; selecting transgenic cloned blastaea in which the EGFP is expressed under a reversed fluorescence microscope, and transplanting the transgenic cloned blastaea into receptor buffalos; and obtaining the transgenic buffalos after full-term pregnancy. After the irradiation by an ultraviolet lamp, the EGFP marker genes are obviously expressed in the heads and limbs of the transgenic buffalo calves, and this proves the method disclosed by the invention can successfully breed transgenic cloned buffalos.
Owner:GUANGXI UNIV
Method for preparing nuleus embryo stem cell from somatic cell transplanting technology
InactiveCN1683524AAvoid ethical issuesAddress immune rejection issuesNew breed animal cellsGenetic material ingredientsGenotypeCell engineering
The method of preparing nucleus embryo stem cell by means of somatic nucleus transplanting technology is one method of somatic nucleus transplanting process to prepare cell. Under microphone operation instrument, nerve nucleus of adult animal is transplanted into denucleated embryo stem cell cytoplast to form embryo stem cell with completely new genome, and the new embryo stem cell is cultured to divide and proliferate to 28 pieces. Embryo stem cell and nerve cell are homologous cells and the prepared cell is nucleus transplanted embryo stem cell with the genome of donor cell. The present invention solves serial problems in somatic nucleus transplanting and can promote the development of cell engineering and tissue engineering relevant to human embryo stem cell.
Owner:SOUTHEAST UNIV
Transgenic vector and transgenic pig for salivary gland tissue-specific expression of exogenous protein and construction method thereof
ActiveCN103695451BImprove playbackSolve efficiency problemsVector-based foreign material introductionAnimal husbandryBiotechnologyPhytase
The invention discloses a transgenic vector of salivary gland tissue specific expression foreign protein and a construction method thereof; the expression vector is constructed by inserting a recombinant fusion gene constructed by glucanase gene, xylanase gene, and phytase gene into a parotid gland-specific transgenic vector; the vector is introduced into pig genome, and transgenic pigs is produced by a somatic cell nucleus transplant technique. The transgenic vector of the invention has hydrolytic action on glucanase in barley raw materials, xylanase in wheat and corn, and phytase in vegetable feed; the pig salivary gland is used as a reactor to realize secretion for the whole life; the addition of the enzyme preparations into feed is replaced permanently, which gives better play to the enzymes; the expression vector of the invention has a piggyback transposon, which greatly increases the transgenosis efficiency, changes the transgenic integration model of common plasmid serial recombination into single locus single copy integration, and thus better simulates the internal environment of biological genes.
Owner:国科润风(广州)生物医药有限责任公司
Preparation method of milk goat mammary gland bioreactor for expressing GP5-M protein in milk
InactiveCN108611371AImmunocompetentAvoid defectsSsRNA viruses positive-senseHydrolasesBiotechnologyAdjuvant
The invention discloses a preparation method of a milk goat mammary gland bioreactor for expressing a GP5-M protein in milk. A gene targeting vector of fusion expression of GP5-M is prepared and thenknocked into a first exon of beta-lactoglobulin of a milk goat to screen target cells; a transgenetic milk goat is prepared by using a somatic nucleus transplantation technology; the mammary gland bioreactor is used for producing milk containing a GP5-M fusion protein; and the pure GP5-M fusion protein is separated from the milk, and an adjuvant is added, so as to prepare a gene engineered subunitvaccine and fulfill the aim of preventing and controlling PRRS.
Owner:NORTHWEST A & F UNIV
Method for obtaining cloned pigs with modified genes through continuous cloning
InactiveCN105755047AOvercome efficiencyOvercome the difficulty of large individual differencesFermentationGenetic engineeringGestationEmbryo
The invention relates to a method for obtaining cloned pigs with modified genes through continuous cloning and belongs to the field of animal reproductive biology.According to the method, tissue fibroblasts of obtained genetically modified pigs serve as donors, donor cells are transferred into enucleated porcine oocytes through the somatic cell nuclear transfer to build a genetically modified pig reconstructed embryos, then electro-fusion and electro-activation are conducted, the donor cells are transferred into estrous surrogate sow fallopian tube expanded portions after being cultured in a pig embryo in-vitro culture solution for 3 h, and after gestation of surrogate sows last for 114 d, a large scale of genetically modified pig individuals are obtained through delivery.The method can solve the problems that in the process that genetically modified individuals are obtained through the gene editing technology, target genetically modified individuals are low in efficiency, and difference between the target genetically modified individuals is large, so that genetically modified cloned pig individuals stable in target gene expression and phenotype can be efficiently produced on a large scale.
Owner:魏红江
Hyperlipemia transgenic miniature pig and preparation method thereof
InactiveCN102086454AReduced activityInhibitory activityFermentationVector-based foreign material introductionFiberHuman apolipoprotein
The invention provides a transgenic miniature pig for preparing a human hyperlipemia disease animal model and a preparation method thereof. A human apolipoprotein CIII expression vector pcDNA-CIII is constructed and expressed by using a gene engineering method; a miniature pig fibroblast capable of expressing the human apolipoprotein CIII is screened out by using cell transfection and cell screening technologies; and the miniature pig is cloned by using a somatic cell nuclear transfer technology. The cloned miniature pig can be used for research into pathogenesis of human hyperlipemia related diseases, and screening and preclinical pretesting of medicine intervention and treatment of related diseases.
Owner:JILIN UNIV
Method for producing parthenogenetic cloned pig by means of continuous cloning technology
ActiveCN103343142ASpeed up development timeImprove controllabilityAnimal reproductionFermentationAnimal scienceNuclear transfer
The invention relates to a method for producing a parthenogenetic cloned pig by means of continuous cloning technology, which belongs to the field of reproductive biology. The method provided by the invention comprises the following steps of: the degreasing of a porcine oocyte, parthenogenetic activation, the separation of a fibroblast, and continuous somatic nuclear transfer; and specifically, the method comprises the following steps of: carrying out the parthenogenetic activation on the degreased mature porcine oocyte to obtain a parthenogenetic embryo, transplanting the parthenogenetic embryo into the body of a surrogate sow to obtain a 25-29 days old living parthenogenetic fetus, establishing a porcine parthenogenetic fetus fibroblast line which is used as a preliminary donor cell, and carrying out continuous cloning by means of somatic cell nuclear transfer technique to obtain a porcine parthenogenetic individual. By adopting the method provided by the invention, the condition that the in-vivo development stops when the porcine parthenogenetic fetus grows for 30 days is overcome, the in-vivo development time of the porcine parthenogenetic fetus is successfully prolonged, and therefore a living parthenogenetic cloned pig can be successfully obtained finally.
Owner:YUNNAN AGRICULTURAL UNIVERSITY
Method for acquiring heterogeneity interspecies-cloned yak premium embryos
InactiveCN103409467AIncrease formation rateIncrease cell numberFermentationGenetic engineeringAgricultural scienceGermplasm
The invention discloses a preparation and in-vitro culture method of interspecies-cloned yak embryos. According to the method, yak-cattle interspecies-cloned embryos are constructed by using yak mammary glandular cells as donor cells and cattle or milch cow enucleated oocytes as receptor cytoplasts by a somatic nucleus transplantation technique; and an electrical activation-chemical activation-reelectrical activation mode and a self-made sequential culture system are utilized to acquire the high-efficiency high-activity yak-cattle interspecies-cloned embryos. The method disclosed by the invention can increase the yak breeding speed, shorten the yak breeding cycle, lower the yak production cost, enhance the breeding efficiency, reduce the damage level of the grassland to some extent, and effectively protect the species resource of the yak. Besides, the invention has important meanings for promoting research of interspecies-cloned yak, transgenic cloned yak embryos and nucleo-cytoplasmic interaction.
Owner:SOUTHWEST UNIVERSITY FOR NATIONALITIES
Preparation method of anti-African swine fever cloned pig
ActiveCN113073114AAcquired ASFV resistanceHydrolasesStable introduction of DNAAfrican swine feverNuclear transfer
The invention discloses a preparation method of an anti-African swine fever cloned pig and belongs to the field of animal genetic engineering and genetic modification. A CRISPR / Cas9 gene knock-in technology is used, a CRISPR / Cas9 targeting vector and a pig Rosa26 gene donor vector are jointly transferred into a pig fibroblast to obtain a positive cell clone, a first intron of the pig Rosa26 gene in the positive clone is knocked in a gene capable of expressing sgRNA-Cas9 of a specific targeted knockout virus pE248R gene, such that the pE248R gene is knocked out in a targeted manner when viruses are propagated in vivo and a pig has ASFV resistance; a cloned embryo is obtained through a somatic cell nuclear transfer technology; and the cloned embryo are transferred into a pig uterus for pregnancy to obtain a cloned pig. The sgRNA-Cas9 system is knocked into a pig body for the first time and stably expressed to obtain ASFV resistance, such that a foundation is laid for breeding research on African swine fever resistance.
Owner:GUANGDONG OCEAN UNIVERSITY
Method for constructing heterogeneous organ transplantation base donor pigs
PendingCN110373390ASpeed up the R&D processFix compatibility issuesGenetic engineeringFermentationAntigenAnimal biotechnology
The invention relates to a method for constructing heterogeneous organ transplantation base donor pigs, and belongs to the field of animal biotechnology. On the basis of fetal fibroblasts, three genes, including GGTA1, CMAH and B4GalNT2, are knocked out by the CRISPR / Cas9 gene editing technology, three humanized modifying genes, including hCD55, hCD59 and hCD46, are inserted at fixed points, and the heterogeneous transplantation base donor pigs modified by CMAHKO / B4GalNT2KO / hCD55 / hCD59 / hCD46 genes is constructed according to a somatic cell nuclear transfer technique. The method solves common problems in heterogeneous organ transplantation, including antigen-antibody immunological rejection reaction and complement regulation, to a certain extent, and lays a foundation for research and development of the specific heterogeneous organ transplantation donor pigs suitable for organs, such as hearts, kidneys and livers as well as tissue such as pancreas islet and skin on the basis of base donor pigs, thereby having high application value and promising application prospect in the construction of the multi-gene modified heterogeneous transplantation donor pigs.
Owner:YUNNAN AGRICULTURAL UNIVERSITY
Reconstructed eggs of albinism model pigs and its construction method and construction method of model pigs
ActiveCN104263754BVector-based foreign material introductionAnimal husbandryDiseaseHypersensitive response
The invention relates to a reconstructed ovum of an albinism model pig and a construction method thereof, and a construction method of the model pig. According to the construction method of the reconstructed ovum, tyrosinase is knocked out to obtain the reconstructed ovum of the albinism model pig. The combination of a CRISPP / Cas9 gene knock-out technique and a somatic nucleus transplantation technique proves that the method for constructing the gene modified pig has high feasibility. The obtained TYR gene knock-out pig has the typical characteristics of albinism, can provide a reliable animal model for researching human albinism, can be applied to toxicity and anaphylaxis detection and many other aspects, and is hopeful to become a standardized experimental animal for albinism-like mice.
Owner:广州中科飞豚生物科技有限责任公司
Who we serve
- R&D Engineer
- R&D Manager
- IP Professional
Why Patsnap Eureka
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com