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170 results about "Nuclear transfer" patented technology

Nuclear transfer is a form of cloning. The steps involve removing the DNA from an oocyte (unfertilised egg), and injecting the nucleus which contains the DNA to be cloned. In rare instances, the newly constructed cell will divide normally, replicating the new DNA while remaining in a pluripotent state. If the cloned cells are placed in the uterus of a female mammal, a cloned organism develops to term in rare instances. This is how Dolly the Sheep and many other species were cloned. Cows are commonly cloned to select those that have the best milk production. On 24 January 2018, two monkey clones were reported to have been created with the technique for the first time.

CICM cells and non-human mammalian embryos prepared by nuclear transfer of a proliferating differentiated cell or its nucleus

InactiveUS6235970B1Simple procedureSimplifying and facilitating procedureNervous disorderMuscular disorderPresent methodNuclear transfer
An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian 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:UNIVERSITY OF MASSACHUSETTS AMHERST

Pluripotent stem cells derived without the use of embryos or fetal tissue

This invention provides a method for deriving precursors to pluripotent non-embryonic stem (P-PNES) and pluripotent non-embryonic stem (PNES) cell lines. The present invention involves nuclear transfer of genetic material from a somatic cell into an enucleated, zona pellucida free human ooplastoid having a reduced amount of total cytoplasm. The present invention provides a new source for obtaining human and other animal pluripotent stem cells. The source utilizes as starting materials an oocyte and a somatic cell as the starting materials but does not require the use, creation and/or destruction of embryos or fetal tissue and does not in any way involve creating a cloned being. The oocyte never becomes fertilized and never develops into an embryo. Rather, portions of the oocyte cytoplasm are extracted and combined with the nuclear material of individual mature somatic cells in a manner that precludes embryo formation. Murine, bovine, and human examples of the procedure are demonstrated. Subsequently, the newly constructed P-PNES cells are cultured in vitro and give rise to PNES cells and cell colonies. Methods are described for culturing the P-PNES cells to yield purified PNES cells which have the ability to differentiate into cells derived from mesoderm, endoderm, and ectoderm germ layers. Methods are described for maintaining and proliferating PNES cells in culture in an undifferentiated state. Methods and results are described for analysis and validation of pluripotency of PNES cells including cell morphology, cell surface markers, pluripotent tumor development in SCID mouse, karyotyping, immortality in in vitro culture.
Owner:STEMA

Cloning pigs using donor nuclei from non-quiescent 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:MASSACHUSETTS UNIV OF

Production of reprogrammed cells with restored potential

InactiveUS20070032447A1Reduce in quantityDifferentiation potential in differentiatedBiocideHydroxy compound active ingredientsNuclear transferGenome
A method for treating cells and / or nuclear transfer units and / or stem cells in culture with such compounds, individually or in combinations, is described. The method results in a globally hypomethylated genome and a restoration of cell differentiation and / or developmental potential, or potentiality. In addition, a method for the in vitro production of reprogrammed cells which have had differentiation potential (totipotential, pluripotential, or multipotential) restored by demethylating the genome is described.
Owner:NUPOTENTIAL INC

Embryonic or stem-like cell lines produced by cross species nuclear transplantation

An improved method of nuclear transfer involving the transplantation of 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.
Owner:UNIV OF MASSACHUSETTS

Method for preparing high muscle content and hypertrophic cardiomyopathy model cloned pig

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

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

Method for generating cloned animals using chromosome shuffling

The present invention concerns the use of chromosomal replacement techniques in the context of producing cloned and transgenic animals, in order to correct chromosome abnormalities or alter autosomal genotypes, and provide for novel breeding pairs by replacing the sex chromosome in animals to be cloned. Replacement of a sex chromosome, or an X or Y chromosome, will result in animals that are autosomally isogenic and sexually non-isogenic (AISN), with "autosomally isogenic" meaning that the paired sets of autosomes (non-sex chromosomes) in each animal are isogenic or identical. Also included in the invention are animals that are both "autosomally" and "allelically" isogenic whereby each particular pair of chromosomes is internally isogenic or identical within a single animal as well as between animals. Such animals are particularly useful in generating a line of cloned mammals using sexual reproduction, without having to undergo nuclear transfer in order to propagate cloned animals.
Owner:ADVANCED CELL TECH INC

Compositions, methods and kits relating to reprogramming adult differentiated cells and production of embryonic stem cell-like cells

The present invention includes compositions, methods and kits for non-nuclear transfer reprogramming an adult differentiated cell obtained from an adult tissue into an ES-like cell. The reprogrammed cell can be converted into an ES-like cell which can be re- or trans-differentiated into various differentiated cell types. The present invention further relates to identification of a novel signaling pathway, and components thereof, which effect reprogramming of a cell. The present invention further comprises compositions, methods and kits for regulating the mammalian cell cycle and cellular proliferation, as well as for treating diseases and for identifying components that affect the cell cycle, and reprogram cells, among other things.
Owner:THE ROCKEFELLER UNIV

CRISPR/Cas9 technology mediated down producing goat VEGF gene fixed-point knock-in method

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

Semi-automatic image cutting method based on nuclear transfer

InactiveCN102663757AEffectively keep the edgeGuaranteed edgeImage analysisFeature setImaging analysis
The invention discloses a semi-automatic image cutting method based on nuclear transfer, which mainly solves the problems that the existing interactive image cutting method can not keep data consistency, the cutting result is not stable, and edges are not clear. The method is realized through the following steps: inputting an image to be cut, obtaining a superpixel set of the input image through a mean shift method, calculating a superpixel color column diagram feature set, building a similarity matrix W through a Bhattacharyya coefficient formula, performing interactive operation on the precut image by a user to obtain a seed superpixel set, building a must-link constrain set M and a cannot-link constrain set C, transferring constrain information M and C to the whole nuclear space through a nuclear transfer method, so as to obtain a nuclear matrix R, clustering the nuclear matrix R through a k mean clustering method, so as to obtain a clustering mark vector, and outputting and displaying a cutting result. The method has the advantages of good data consistency, stable cutting result, simplicity of operation and clear edges, and can be applied to image retrieval, the technology of converting 2D to 3D, target detection and tracking, and medical image analysis.
Owner:XIDIAN UNIV

Microelectromechanical devices useful for manipulating cells or embryos, kits thereof, method of making same, and methods of use thereof

The present invention relates generally to microelectromechanical systems (MEMS) devices for the manipulation of cells or groups of cells, such as oocytes, embryos, and sperm. In particular, the present invention relates to Cell Labeling MEMS devices (2F), Microinjection MEMS devices, IntraCytoplasmic Sperm Injection (“ICSI”) MEMS devices, Zona Coring MEMS devices, Enucleation MEMS devices, Enucleation / Nuclear Transfer MEMS devices, and Cytoplasmic Transfer MEMS devices. The present invention also relates to kits containing the MEMS devices of the present invention.
Owner:MATHEWS SHEPHERD MCKAY & BRUNEAU P A

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

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:ADVANCED CELL TECH INC

Nuclear transfer with porcine embryonic stem cells

Provided is a method for the production of transgenic animals, especially pigs, by the use of nuclear transfer from genetically modified or other embryonic stem cells to either enucleated oocytes which were matured in vivo or in vitro and activated or to enucleated zygotes.
Owner:BOARD OF TRUSTEES OF THE UNIV OF ILLINOI THE

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

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

Process of making transgenic mammals that produce exogenous proteins in milk and transgenic mammals produced thereby

The invention relates to a method of producing a protein of interest, comprising making a non-human transgenic mammal that produces said protein in its milk, obtaining said milk from the non-human transgenic mammal and purifying said protein of interest from the milk. Transgenic bovine animals were generated, which are able to produce human growth hormone in mammary glands. The method involves cloning of a genetic construct encoding hGH gene and beta casein promoter conveniently in an expression vector. It also includes transfection procedures into fetal bovine somatic cells, generally fibroblasts, and the nuclear transfer into enucleated bovine oocytes, generating thus transgenic embryos. The method also includes other procedures to generate transgenic embryos for the further expansion of the transgenic herd, such as the subcloning of transgenic female bovines, the superovulation of transgenic cows and their insemination with semen from a non-transgenic or a transgenic male bovine, and the superovulation of non-transgenic cows and their insemination with semen from a transgenic male bovine. Afterwards, transgenic embryos give rise to transgenic cattle that produce human growth hormone in huge amounts in their milk, from which the hormone is completely purified and analysed to fulfill all the requirements for the manufacture of a pure biopharmaceutical product.
Owner:STERRENBELD BIOTECH NORTH AMERICA

Nuclear transfer nuclei from histone hypomethylated donor cells

The present invention provides a method of producing an animal embryo, the method comprising transferring from a nuclear donor cell which has been selected on the basis that it is histone hypomethylated at least a portion of the nuclear contents including at least the minimum chromosomal material able to support development into a suitable recipient cell.
Owner:MEDICAL RESEARCH COUNCIL

Method of constructing nuclear-transplanted egg parthenogenetic embryo and parthenogenetic mammal

The present invention is to provide a method of constructing a nucleus-implanted egg, a parthenogenetic embryo and a parthenogenetic mammal each having 2 haploid genome sets originating in mammarian ova, and provides methods of constructing a nucleus-implanted egg having a haploid genome set derived from ng ovum and a haploid genome set from fg ovum, a parthenogenetic embryo and a parthenogenetic mammal, which includes the steps of (1) introducing a primitive ovarian follicle egg (ng ovum) into a nucleus-deleted egg in a germinal vesicle stage (GV stage egg) and then developing them to MII phase (second meiosis metaphase) by in vitro maturing and culturing to prepare a first nucleus-implanted egg, and (2) extracting MII phase chromosome from said first nucleus-implanted egg and introducing it into other MII phase egg (fg ovum) to prepare a second nucleus-implanted egg, wherein ovum from which an imprinted gene that undergoes gene modification posteriori during the generation of sperm is deleted is used as the ng ovum or fg ovum.
Owner:TOKYO UNIVERSITY OF AGRICULTURE

A method for processing bovine somatic cell cloned embryos based on somatic cell nuclear transfer

The invention discloses a method for processing bovine somatic cell cloned embryos constructed on the basis of somatic cell nuclear transplantation, which comprises the following steps of: injecting bovine somatic cells to be transplanted into denucleated oocyte to carry out electrofusion; and an hour later after the electrofusion is completed, selecting the successfully fused bovine somatic cellcloned embryos and processing the bovine somatic cell cloned embryos for 12 hours by 1muM of Oxamflatin. By the method, the developmental rate and the quality of the bovine somatic cell cloned embryos can be obviously improved and the bovine somatic cell cloned embryos can be efficiently produced in vitro.
Owner:NORTHWEST A & F UNIV

Cloning pigs using non-quiescent differentiated donor cells or nuclei

InactiveUS7291764B1Suppress pro-inflammatory medicationHigh expressionTissue cultureFermentationBiotechnologyPresent method
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:UNIVERSITY OF MASSACHUSETTS AMHERST

Method for breeding transgenic livestock rich in polyunsaturated fatty acid

The invention discloses a method for breeding transgenic livestock rich in polyunsaturated fatty acid. The method comprises the following steps of: guiding DNA molecules which have a sequence 1 and a sequence 2 in a sequence list into somatic cells of a mammal to obtain transgenic cells in which a delta-12 fatty acid dehydrogenase and an omega-3 fatty acid dehydrogenase are expressed; cloning embryos by using the transgenic cells as nuclear transfer donor cells and using isolated ovocytes as nuclear transfer acceptor cells by nuclear transfer technology; and transplanting the cloned embryos into a uterus of the livestock by a non-operation method for gestation to obtain the transgenic livestock. The method of the invention can ensure that the delta-12 fatty acid dehydrogenase and the omega-3 fatty acid dehydrogenase are stably expressed in the body of a transgenic pig and that the content of omega-3 polyunsaturated fatty acids (PUFAs) in meat of the livestock is improved obviously.
Owner:BEIJING GEFUCURE BIOTECHNOLOGY LIMITED COMPANY

Method for producing pluripotent stem cells

Methods are provided for producing a human embryo capable of developing to the blastocyst stage. The method includes transferring a human somatic cell genome into a mature human oocyte by nuclear transfer and activating the oocyte, without removing the oocyte genome. Pluripotent human embryonic stem cells, and methods of obtaining these, are also provided.
Owner:NEW YORK STEM CELL FOUND

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
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