Patents
Literature
Hiro is an intelligent assistant for R&D personnel, combined with Patent DNA, to facilitate innovative research.
Hiro

169 results about "Nuclear transplantation" patented technology

Nuclear transplantation is a method in which the nucleus of a donor cell is relocated to a target cell that has had its nucleus removed (enucleated). Nuclear transplantation has allowed experimental embryologists to manipulate the development of an organism and to study the potential of the nucleus to direct development.

Overexpression porcine co-stimulatory 4-1BB vector and application thereof

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

Stem cell-derived endothelial cells modified to disrupt tumor angiogenesis

InactiveUS20060024280A1Enhance the ability of the endothelial cells to disrupt and inhibit tumor angiogenesisHigh sensitivityBiocideGenetic material ingredientsAbnormal tissue growthTumor angiogenesis
The present invention provides cloned, genetically modified, endothelial cells, and the stem cells from which they are derived, which are produced by somatic cell nuclear transfer. The invention further provide novel therapeutic methods in which such cells are administered to a patient with tumors to inhibit and / or disrupt angiogenesis of the tumors, thereby inhibiting tumor growth and killing tumor cells.
Owner:ADVANCED CELL TECH INC

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

Embryonic or stem-like cell lines produced by cross species nuclear transplantation and methods for enhancing embryonic development by genetic alteration of donor cells or by tissue culture conditions

An improved method of nuclear transfer involving the transplantation of differentiated donor cell nuclei into enucleated oocytes of a species different from the donor cell is provided. The resultant nuclear transfer units are useful for the production of isogenic embryonic stem cells, in particular human isogenic embryonic or stem cells. These embryonic or stem-like cells are useful for producing desired differentiated cells and for introduction, removal or modification, of desired genes, e.g., at specific sites of the genome of such cells by homologous recombination. These cells, which may contain a heterologous gene, are especially useful in cell transplantation therapies and for in vitro study of cell differentiation. Also, methods for improving nuclear transfer efficiency by genetically altering donor cells to inhibit apoptosis, select for a specific cell cycle and / or enhance embryonic growth and development are provided.
Owner:ADVANCED CELL TECH INC

Efficient Somatic Cell Nuclear Transfer In Fish

The present disclosure provides methods of producing enucleated cells by photoablation. Such enucleated cells may be used as recipient cells for Somatic Cell Nuclear Transfer and cloning. The nuclear donor and / or enucleated recipient cells may be any fish cells, such as zebrafish, koi, or medaka fish cells. Such methods may be used to efficiently produce transgenic fish including by way of example zebrafish, koi, and medaka fish.
Owner:MICHIGAN STATE UNIV

Method for generating immune-compatible cells and tissues using nuclear transfer techniques

InactiveUS20020046410A1Easily propagatableEasily transfectedCompounds screening/testingNew breed animal cellsNuclear transferTransgene
This invention relates to methods for making immune compatible tissues and cells for the purpose of transplantation and tissue engineering, using the techniques of nuclear transfer and cloning. Also encompassed are methods for determining the effect on immune compatibility of expressed transgenes and other genetic manipulations of the engineered cells and tissues.
Owner:LANZA ROBERT +1

Method for in vitro incubation of macrobrachium nipponense

The invention relates to a method for in vitro incubation of fertilized eggs of macrobrachium nipponense, which is characterized in that the method comprises the steps of selection of parent shrimps, mating and oviposition, treatment of incubation water, egg stripping, egg washing, incubation, daily management and the like. The method for the in vitro incubation of the fertilized eggs of the macrobrachium nipponense solves the problem of biological characteristics that the fertilized eggs depend on parent bodies to be incubated, lays a foundation for genetic operations of the macrobrachium nipponense such as gene transfer, nuclear transplantation and the like, fills the research gap of the field, and has important practical application value.
Owner:FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI

Use of RNA interference for the creation of lineage specific ES and other undifferentiated cells and production of differentiated cells in vitro by co-culture

Methods for making human ES cells and human differentiated cells and tissues for transplantation are described, whereby the cells and tissues are created following somatic cell nuclear transfer. The nuclear transfer donor is genetically modified prior to nuclear transfer such that cells of at least one developmental lineage are de-differentiated, i.e., unable to develop, thereby resolving the ethical dilemmas involved in reprogramming somatic cells back to the embryonic stage. The method concomitantly directs differentiation such that the desired cells and tissues may be more readily isolated.
Owner:ADVANCED CELL TECH INC

Method for improving pig cloning efficiency on basis of inhibition of H3K9me3 methylation

The invention discloses a method for improving the pig cloning efficiency on the basis of inhibition of H3K9me3 methylation. According to the method, H3K9me3 methylation is inhibited by inhibiting expression of Suv39H1 and Suv39H2 genes in nuclear donor cells during nuclear transplantation of a cloned embryo of a pig and overexpression of KDM4A mRNA during nuclear transplantation of a reconstructed embryo, and the pig cloning efficiency is further improved. Specifically, according to the method for improving the pig cloning efficiency, effective siRNA resisting Suv39H1 and Suv39H2 genes is co-transfected in the nuclear donor cells before nuclear transplantation, overexpression of the Suv39H1 and Suv39H2 genes in the nuclear donor cells is inhibited, accordingly, the high methylation level of H3K9me3 in the nuclear donor cells is inhibited, the processed nuclear donor cells are subjected to nuclear transplantation, cytoplasm microinjection of KDM4A mRNA is performed after the reconstructed embryo obtained after nuclear transplantation is activated, methylation of histone H3K9me3 in the reconstructed embryo is further inhibited, and the cloning efficiency of the pig is finally improved.
Owner:SOUTH CHINA AGRI UNIV

Nuclear transfer embryo formation method

A nuclear transfer embryo is formed by destabilizing microtubules of an oocyte, whereby essentially all endogenous chromatin collects at a second polar body during meiosis of an oocyte. The oocyte is fused with the nucleus of a donor somatic cell of the same species of said oocyte prior to cessation of extrusion of the second polar body from the oocyte, thereby forming the nuclear transfer embryo. In one embodiment, the nuclear transfer embryo is employed to impregnate an animal, such as a mammal. In another embodiment, the donor nucleus is transgenic.
Owner:TRUSTEES OF TUFTS COLLEGE

Bank of stem cells for producing cells for transplantation having HLA antigens matching those of transplant recipients, and methods for making and using such a stem cell bank

Methods for producing stem cell banks, preferably human, which optionally may be transgenic, e.g., comprised of homozygous MHC allele cell lines are provided. These cells are produced preferably from parthenogenic, IVF, or same-species or cross-species nuclear transfer embryos or by dedifferentiation of somatic cells by cytoplasm transfer. Methods for using these stem cell banks for producing stem and differentiated cells for therapy, especially acute therapies, and for screening for drugs for disease treatment are also provided.
Owner:ADVANCED CELL TECH INC

Preparation and selection of donor cells for nuclear transplantation

The invention relates to a method of synchronizing a population of somatic cells in G1 for purposes of preparing the cells for nuclear transfer or nuclear transplantation by using mechanical shake-off and selection of mitotic doublet cells. This method may further comprise cooling of the cells or other means of synchronizing the cells in G1 phase for longer periods of time. The invention also relates to the use of a synchronized population of rapidly, dividing somatic cells obtained by these methods as a source of donor nuclei or chromatin for use in nuclear transfer or nuclear transplantation.
Owner:UNIV OF MASSACHUSETTS A PUBLIC INSTION OF HIGHER EDUCATION BY THE COMMOMWEALTH OF MASSACHUSETTS AS REPRESENTED BY ITS AMHERST CAMPUS

Cloning pigs using donor nuclei from differentiated cells

An improved method of nuclear transfer involving the transplantation of donor differentiated pig cell nuclei into enucleated pig oocytes is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring. Production of genetically engineered or transgenic pig embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
Owner:UNIV OF MASSACHUSETTS

Bovine oocyte in vitro maturation culture solution and culture method

The invention provides a bovine oocyte in vitro maturation culture solution and a culture method. The maturation culture solution consists of 10% of fetal calf serum and 1% of ITS-G, as well as 0.075IU / mL of HMG, 1[mu]g / mL of 17[beta]-estradiol, 10ng / mL of EGF, 2.2mg / mL of bFGF and 50ng / mL of CXCL12. The culture solution, when applied to bovine oocyte in vitro maturation culture, can improve maturation quality of bovine oocyte in vitro culture, a development rate of in vitro fertilized embryos as well as a development rate of somatic cell nuclear transplantation embryos and blastocyst quality.
Owner:NORTHWEST A & F UNIV

Method and device for transplantation of femtosecond laser nucleus

The invention relates to a method and equipment for femtosecond laser somatic cell nuclear transplantation, in particular to a micro-nano control method which adopts a light knife effect induced by femtosecond laser two-photon and optical tweezers gradient force induced by high photon density. A pump laser is started-up; the laser produced by the pump laser is introduced into a resonator of a femtosecond laser; when the oscillation is done, the energy of the ultrashort femtosecond laser pulse is amplified through a regenerative amplifier, and the pulse number and the repeated frequency of laser output are chosen. The emitted femtosecond laser is divided into two beams of light, one of which used for realizing the femtosecond optical tweezers, and the other of which is used for realizing the function of the femtosecond optical tweezers through a half-permeable and semi-reflecting mirror, and then the two beams of light are coupled with an optical path through another half-permeable andsemi-reflecting mirror to realize the amplification of a spot light area; then the light beams is focused on a target cell, and the harmless transplantation operation of the somatic cell nuclear is done. A switch for controlling a light valve is arranged in a software control system, which can not only realize the operation of the laser spot on the target cell, but also adjust the scanning speed and change the exposure time to meet different requirements and enhance the success rate of the somatic cell nuclear transplantation.
Owner:JIANGSU UNIV

New method for gene injection for somatic cell nuclear transfer reconstructed embryo

InactiveCN106520838AMeet biosafety requirementsAvoid screening difficultiesMicroinjection basedFermentationLarge fragmentEmbryo transfer
The invention discloses a new method for gene injection for a somatic cell nuclear transfer reconstructed embryo. The method comprises the following steps: target gene preparation, somatic cell nuclear transfer and a microinjection technology. An expression vector is constructed after a target gene is obtained, the cell nucleus of an oocyte is removed by virtue of micromanipulation, then a somatic cell is injected in the space of the denucleated oocyte, then an exogenous gene is directly injected in the somatic cell, the reconstructed embryo is constructed through one-step electric fusion and activation, and the embryo is transferred to obtain a transgenic progeny. According to the method disclosed by the invention, large-fragment cell gene transfection and screening are avoided, the transgenosis efficiency is increased, the time is saved, and the transgenosis application range is expanded. Importantly, the somatic cell is injected in a transpanent zone during a nuclear transfer operation process, thus microinjection for the exogenous target gene is facilitated.
Owner:INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI

Cloning using rapidly matured oocytes

A method of producing a cloned or genetically modified non-human mammalian embryo comprising: (a) providing a cell culture comprising a plurality of in vitro matured oocytes; (b) preferentially selecting from the cell culture a rapidly matured oocyte or developmentally competent oocyte; (c) transferring DNA from a donor cell derived from non-human mammalian tissue to the matured oocyte to form a nuclear transfer unit; and (d) culturing said nuclear transfer unit to form an embryo. At the initiation of maturation, oocytes are preferably beyond the GV-II stage of prophase I. Porcine oocytes most preferably mature in about 20-28 hours.
Owner:UNIV OF GEORGIA RES FOUND INC

Rearing process for Shuangji cloned pig by gene engineering technology

A genetic engineering technology for preparing the clonal pig by gene knock-out and somatic cell nucleus transplantation includes such steps as homoglous recombining to destroy the muscle growth inhibin gene of somatic cell, externally screening the targeted cells, and nucleus transplanting. Its advantage is stable inheritance of excellent characteristics.
Owner:INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE

Method for cloning animals with targetted genetic alterations by transfer of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells

An improved method of nuclear transfer employing long-term cultured somatic cells as the donor cells and enucleated oocytes as the recipient cells to produce dividing cybrids. Such cybrids are useful for developing viable animals clones when nurtured in a suitable host environment.
Owner:GOVERNOR OF KAGOSHIMA PREFECTURE THE +1

Mammal body-cell neucleus transplanting method

InactiveCN1814750AImprove reprogramming abilityImprove developmentTissue cultureFermentationMammalF1 generation
This invention provides a nucleus transplantation method for mammal somatic cells characterizing in taking the oocyte of cross-fertilized F1 generation of the mammal as the recipient somatic cell for nucleus transplantation, which can increase the re-programming ability of recipient cytoplast of nucleus transplantation of mammals obviously, improve the growth of re-structured embryo to increase the clone efficiency of somatic cells.
Owner:SHANGHAI JIAO TONG UNIV AFFILIATED CHILDRENS HOSPITAL +1

Transgene pig for preparing animal model for human diseases and culturing method thereof

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

ActiveCN105177044ABigHigh blood circulationAnimal reproductionEmbryonic cellsDiseaseEmbryo
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

Method for the rapid selection of homozygous primary cell lines for the production of transgenic animals by somatic cell nuclear transfer

The present invention provides for the production of homozygous primary cells that carry a specific transgenic integration of interest on both chromosomes by bypassing breeding. These cell lines can they be used for the accelerated production of homozygous transgenic animals by somatic cell nuclear transfer. The invention is thus useful in the production of transgenic ungulate animals capable of producing desired biopharmaceuticals in their milk at higher yield than a comparable heterzygote. By combining the selection techniques of the current invention with somatic cell nuclear transfer it can be applied to large animals, where there is a strong need to shorten the time to homozygosity.
Owner:GTC BIOTHERAPEUTICS INC

Cloning of transgenic animals comprising artificial chromosomes

The invention is directed in part to totipotent cells that have one or more artificial chromosomes; processes for producing such cells; processes for using such cells (e.g., nuclear transfer); transgenic embryos and transgenic animals cloned from such cells; and processes for producing such embryos and animals.
Owner:SAB LLC

Artificially induced tetraploid crucian cell line and culture method and application thereof

The invention discloses an artificially induced tetraploid crucian cell line which is preserved in the China General Microbiological Culture Collection Center with the preservation number of CGMCC No.9155. The culture method comprises the following steps: by using in vitro cultured diploid crucian cells as a raw material, treating by SP600125 when the cell growth density reaches the degree of fusion being 80-90%; then culturing by a conventional culture medium without SP600125; after culturing a generation, carrying out flow analysis; detecting the content of DNA (Deoxyribonucleic Acid); meanwhile, detecting the chromosome ploidy of the cultured crucian cell; and artificially inducing and sorting and purifying to obtain the in vitro induced tetraploid crucian cell line. The cell nucleus of the tetraploid crucian cell line disclosed by the invention is transplanted to a denucleated egg by using an induced pluripotent stem cell method or adopting a method combining nuclear transplantation. The artificially induced tetraploid crucian cell line can be used for quickly and efficiently creating tetraploid crucian.
Owner:HUNAN NORMAL UNIVERSITY

Method for producing transgenic pigs through overexpression HOXA10 genes

The invention belongs to the technical field of pig transgenosis and particularly relates to a method for producing transgenic pigs through overexpression HOXA10 genes. The method is characterized in that constructed HOXA10 gene expression carriers are subjected to lipofection and sulfate (G418) screening to obtain HOXA10 transgenic cell lines with stable expression; the HOXA10 transgenic cell lines are transplanted into denucleated pig oocytes with HOXA10 transgenic pig fibroblasts as nuclear donors through an artificial nuclear transplantation method to form reconstructed embryos; the reconstructed embryos are transplanted into a receptor sow oviduct, and after a sow gives birth to the HOXA10 transgenic pigs and molecular identification is conducted, masculine HOXA10 transgenic pigs are screened out. The transgenic pigs obtained through the method provide materials for research about the mechanism of improving the reproductive capacity through the HOXA10 genes and provide an animal model for revealing other functions of the HOXA10 genes.
Owner:HUAZHONG AGRI UNIV

Method for improving pig meat quality

The invention belongs to the technical field of animal genetic engineering and particularly relates to a method for improving pig meat quality. The method is characterized in that a peroxisome proliferator-activated receptor (PPAR) gamma gene serves as an important candidate gene for improving the meat quality, a fibroblast cell line of a 30-day-old fetus of a large white pig is established, a SwaI linearized pN1-MCK-PPAR gamma2 expression vector is shifted to the 30-day-old fetus fibroblast cell line of the large white pig through an electrotransfection method, and a PPAR gamma transgenic pig is prepared in a method of somatic nucleus transplantation. The influence of muscle tissue overexpression PPAR gamma genes on meat traits such as intramuscular fat deposition is verified in a transgenic pig, the contradiction of simultaneously selecting meat quality and meat quantity in conventional animal breeding is overcome, and the novel method is provided for cultivating lean meat pigs with good meat quality.
Owner:HUAZHONG AGRI UNIV
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
PatSnap group products