35 results about "LIN28" patented technology

The invention discloses a novel ips cell inducing method and provides application of inducing genes Oct3/4, Klf4, Sox2, P53, L-Myc, Lin28, RARg and Lrh-1 in inducing isolated mammal somatic cells to generate iPS multipotent stem cells. By an iPS cell establishing method, a method for inducing human iPS stem cells free of exogenous serum, trophoblast cells and animal source is established on the basis of no insertion. The method shortens reprogramming time of the iPS multipotent stem cells and improves cloning generation efficiency, and obtained iPS cells are higher in dryness.

The present invention concerns the delivery of certain reprogramming factor proteins into cells, such as differenti-atedsomatic cells, in order to induce the epi-genetic reprogramming of the cell so it becomes a pluripotent stem cell. The reprogramming factor protein(s) may be Sox2, Klf4, Oct3/4, c-Myc, Lin28, Nanog, or any protein with reprogramming (-enhancing) activity. These proteins may be linked recombinantly or chemically to a cell penetrating peptide that helps facilitate the introduction of these proteins into the target cell and may be preferably expressed in mammalian cells to maintain them in active forms. Accordingly, the present method of inducing pluripotent stem cell (iPS) formation avoids the use of viral or DNA-based expression vectors or the expression of reprogramming factor genes within target cells, which are known to be harmful to the host target cell and cause cancer.

The invention relates to a hoofed mammal inducible multipotential stem cell and a preparation method thereof. The method comprises the following steps of: A) constructing an expression vector carrying a transcription factor, wherein the transcription factor is Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) introducing the transcription factor in the step A) into the cells of the hoofed mammal in a combining form; picking clones of which the form is similar to that of the embryonic stem cell for subculturing; and screening the cell clones meeting the characteristic of the embryonic stem cell to obtain the hoofed mammal iPS cell. The method contributes to determining the most proper culturing condition and the most proper culturing method for establishing an ES cell line of pig, sheep, cattle and other hoofed mammals; and experimental models for various genetic diseases of human beings can be established with the hoofed mammal iPS cell line.

The invention relates to a method for reprogramming cells from aged donors or senescent cells to pluripotent cells that have lost marks of senescence. In particular, the invention relates to an ex vivo method for preparing induced pluripotent stem cells (iPSCs) from a target cell population comprising cells from aged donors or senescent cells, said method comprising the steps of culturing said target cell population under appropriate conditions for reprogramming said cells into iPSCs, wherein said appropriate conditions comprises increasing expression in said target cells, of at least the following reprogramming factors: Oct4, Klf4, Sox2, c-Myc, Lin28 and, optionally Nanog.

The invention relates to interference small molecule ribose nucleic acid (RNA) of a targeted LIN28 gene and glioma migration resistant application thereof. The interference small molecule RNA is in a base sequence shown as SEQ NO.1. In the invention, the interference small molecule RNA of the gene with the effect of inhibiting glioma proliferation is provided, a glioma cell model is used for researching the inhibiting effect of the interference small molecule RNA on the glioma proliferation, and a new method is provided for genetic therapy of glioma.

The present invention relates to the production of avian induced pluripotent stem cells from non-pluripotent somatic cells, including embryonic fibroblasts and adult somatic cells. In this method, avian (including quail or chicken) somatic cells are reprogrammed into a state closely resembling embryonic stem cells including the expression of key stem cell markers alkaline phosphatase, etc. by transfecting/transducing the non-stem cells with genes (preferably using a non-integrating vector as otherwise described herein or alternatively an integrating vector, such a lentiviral vector, retroviral vector or inducible lentiviral vector, among others) which express at least nanog, Lin28 and cMyc. In preferred aspects of the invention, the transfected/transduced vectors express nanog, Lin28, cMyc, Oct 4 (POU5F1 or PouV), SOX2 and KLF4. The induced stem cells which are produced contribute to all 3 germ layers, the trophectoderm and in certain aspects, the gonad in chimeric offspring.

The present invention provides an induced cancer cell capable of self-replication in vitro which is useful in cancer therapy research and the research for cancer-related drug discovery, processes for production thereof, cancer cells induced by the malignant cells, and applications of these cells.

The present invention provides an induced cancer stem cell capable of proliferation (self-replication) in vitro, wherein the induced cancer stem cell has the following two characteristics:

• (1) expressing the six genes (self-renewal related genes) POU5F1, NANOG, SOX2, ZFP42, LIN28, and TERT selected from a certain group of genes; and
• (2) having an aberration which is either (a) a mutation in an endogenous tumor suppressor gene or (b) increased expression of an endogenous cancer-related gene.

The invention relates to a preparation method of an inducible pluripotent stem cell of a goat. The preparation method comprises the following steps: A) constructing a lentiviral vector carrying a transcription factor which is selected from Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) infecting a goat adult cell by combining the transcription factor with the lentiviral vector prepared in the step A, selecting clone with a shape similar to an embryonic stem cell for subculturing, and preparing the inducible pluripotent stem cell of the goat by screening cell clone in accordance with the characteristics of the embryonic stem cell. According to the preparation method, an optimal culture condition and method of a goat ES (embryonic stem) cell line construction can be established; the inducible pluripotent stem cell of the goat is an excellent vector for goat gene targeting, and the inducible pluripotent stem cell of the goat facilitates the revelation of each gene function and the complex development of the goat.

The invention relates to a sheep induced pluripotent stem cell and a preparation method thereof. The preparation method comprises the steps of: A) constructing lentivirus vectors with transcription factors, wherein the transcription factors are selected from Oct4, Sox2, c-Myc, K1f4, Lin28, Nanog, SV40 and hTERT; and B) infecting sheep adult cells to the transcription factors in a combining form by adopting the lentivirus vectors obtained in the step A), selecting cloned passages with forms similar to those of embryonic stem cells for culturing, and sieving the cells satisfied with characteristics of embryonic stem cells for cloning so as to obtain the sheep induced pluripotent stem cells. The sheep induced pluripotent stem cell and the preparation method thereof, disclosed by the invention, are good for determining most suitable culture condition and method of sheep ES cell establishment system; the sheep induced pluripotent stem cell is a good carrier for sheep gene targeting; and the sheep induced pluripotent stem cell is beneficial to revealing gene functions and complex development events of sheep and can be used for improving species and promoting economic growth.

Expression of reprogramming factors such as Sox2, klf4, c-myc, Nanog, LIN28 and Oct4 followed by culture in a MEK inhibitor and a GSK3 inhibitor reprograms tissue cells. The invention provides new uses of these inhibitors, for example in inducing completion of the transcriptional resetting of so-called pre-pluripotent (pre-iPS) stem cells, for example as obtained from mammalian neural stem cells or epiblast stem cells treated with single or combinations of the reprogramming factors, expressed transiently or by integrative vectors. Also provided are systems for reprogramming an epiplast stem cells independently of the use of there inhibitors.

The invention relates to the technical field of medical biological detection and provides novel uses of WDR1, NMI and LIN28 proteins, particularly an application in preparation of a kit for assessingbrain glioblastoma prognosis. The invention further provides a kit and detection method performing brain glioblastoma prognosis assessing through quantitative detection of the WDR1, NMI and LIN28 proteins by adopting an immunohistochemical method. Expression levels of the WDR1, NMI and LIN28 proteins in brain glioblastoma tissues are measured by utilizing an immunohistochemical technique and system scoring, and it is determined that the expression levels of the WDR1, NMI and LIN28 proteins are related with prognosis of patients with brain glioblastoma after operations by combination with postoperative follow-up observation information. The combination of the WDR1, NMI and LIN28 proteins can be used for preparing a protein molecular marker for determining prognosis of patients with brain glioblastoma, and is of great directive significance for brain glioblastoma molecular subtyping, postoperative monitoring of patients and sequential therapy.

The invention provides a method for reprogramming an immortalized lymphocyte line into induced pluripotent stem cells. The reprogramming method mainly comprises the step of introducing additional plasmids for expressing exogenous reprogramming factors into the immortalized lymphocytes through a nuclear transformation method, wherein the reprogramming factors comprise OCT3/4, shP53, SOX2, KLF4, LIN28 and L-MYC. According to the method, the immortalized lymphocyte line can be efficiently reprogrammed into the iPSC, and the additional plasmids cannot be integrated into the genome of host cells and can be gradually lost along with cell proliferation, so that exogenous expression can be timely silenced, and the cell tumorigenesis risk is reduced. The obtained iPSC has typical stem cell morphology and characteristics similar to ESC, expresses cell stemness genes, and has differentiation potential of differentiating into ectoderm, mesoderm and endoderm tissues. The method is high in reprogramming efficiency and low in tumorigenicity, the clinical research value of the immortalized lymphocyte line is expanded, and the method has application prospects.

A method of preparing a population of iPS cells including (i) expressing one or more Yamanaka factors selected from Oct3/4, Sox2, Klf4, Myc, Nanog and Lin28, and reducing the amount and/or activity of Menin (Men1) in a population of target cells, and (ii) optionally isolating the iPS cells from the target cell population; and a method of enhanced differentiation of a first cell into a somatic cell of a tissue of interest, including (i) treating a cell with a differentiation factor of the tissue of interest, and (ii) reducing the amount and/or activity of Menin (Men1) in a population of target cells.

A major object of the present invention is to provide a method for producing induced pluripotent stem cells with low tumorigenesis potential and high induction efficiency.

The invention provides a method for producing induced pluripotent stem cells comprising the step of introducing one or more nucleic acids that facilitate expression of at least one gene selected from the group consisting of NANOG, SOX2, OCT3/4, KLF4, LIN28, and c-MYC into somatic cells.

The invention relates to a method for efficiently producing a cloned chicken. The method comprises the following steps: (1) constructing a lentivirus overexpression vector of four factors, namely OCT4, SOX2, NANOG and LIN28, and transfecting a black-feather Langshan chicken fibroblast CEF; (2) inducing the Langshan chicken fibroblast CEF transfected with the OSNL into iPSCs (induced pluripotent stem cells) by utilizing an induction culture medium, and carrying out passage and identification on the iPSCs formed by induction; (3) performing induced differentiation on the iPSCs formed by induction by utilizing an induction culture medium to obtain PGC-like, and identifying the PGC-like; (4) collecting PGC-like cells, injecting the PGC-like cells into recessive white feather chick embryo blood vessels which are hatched for 2.5 days by a chick embryo blood vessel injection method, and determining that the PGC-like cells can migrate and homing in the receptor recessive white feather chick by a frozen section method; and (5) continuously hatching the recessive white feather receptor chick embryos injected with the PGC-like cells of the black-feather Langshan chickens until the recessive white feather receptor chick embryos are hatched. The microsatellite technology proves that the black feather offspring is derived from the PGC-like cell of the donor. According to the invention, a large number of cloned chickens having the same genetic characteristics as donors can be produced.

The present invention relates to a method for production of a cell population containing a pluripotent stem cell, said method comprising a step of treating a somatic cell which has been contacted with nuclear reprogramming factors under nutrient-starved condition, and/or a step of treating the somatic cell with an agent capable of arresting cell cycle. The present invention allows induction and growth of pluripotent stem cells at high frequency, and it also allows production of pluripotent stem cells with high efficiency. The nuclear reprogramming factors to be used may be any selected from the group consisting of OCT4, SOX2, c-MYC, KLF4, NANOG and LIN28.

The invention discloses hsa-let-7 familial miRNA and applications of target genes of the hsa-let-7 familial miRNA in diagnosis and treatment for EBOV infection. The hsa-let-7 familial miRNA disclosed by the invention is hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p, the target genes of the hsa-let-7 familial miRNA are MAB21L3 gene, MEIS3 gene, DPP6 gene, IGDCC3 gene, CHRD gene, FASLG gene, LIN28 gene, E2F5 gene, IL-6 gene and E2F6 gene. The hsa-let-7 familial miRNA and the target genes of the hsa-let-7 familial miRNA can be taken as biological molecular markers for diagnosing EBOV infection, and also can be taken as biological target molecules for the design of the medicines capable of resisting EBOV infection.

The invention provides a novel method for reprogramming human amniotic epithelial cells into induced pluripotent stem cells, comprises following steps: introducing a plurality of oriP/EBNA1 free plasmid vectors which carrying six reprogramming factors including five transgenes such as OCT4, SOX2, KLF4, LIN28 and L-MYC and one P53 gene shRNA into hAECs by using a non-lipid cation transfection reagent NeofaectTM. The method is simple in process, convenient to operate and low in cost, and the free plasmid vector is a non-gene integrated reprogramming method, so that tumorigenicity is reduced; thewhole reprogramming process is short in time consumption and high in efficiency; the obtained hAE-iPSCs clone is subjected to enlarged culture in a TeSRTM-E8TM-human recombinant vitronectin system without animal origin, which lays a foundation for clinical application.

The invention discloses a production method for buffalo induced pluripotent stem cells. Specific transcription factors such as Oct4, Sox2, Klf4, c-Myc, Nanog and Lin28 and a cell immortal gene SV40large T (LT) are carried by retrovirus vectors, and buffalo somatic cells such as fetus fibroblasts are directly reprogrammed into the induced pluripotent stem cells with characteristics similar to those of embryonic stem cells. Due to the generation of the buffalo induced pluripotent stem cells, a new method for obtaining buffalo stem cells can be provided, the problems that the buffalo embryonic stem cells are difficult to separate and cultivate, and cell lines are difficult to establish are solved to some extent, moreover, sources of stem cells are provided for the research of signal paths of the buffalo stem cells and the establishment of the cell lines, and a new method is provided for directional genetic improvement of buffalos at the same time.

The invention discloses a multifunctional intimal stem cell hematopoietic molecular material and a method, relates to the field of biological pharmacy, and provides the following scheme aiming at the problems of low cell differentiation induction efficiency, poor cell functionality and the like in the prior art: the multifunctional intimal stem cell hematopoietic molecular material comprises a pluripotent hematopoietic stem cell, a specific gene, a somatic cell, a niche, a lentiviral vector and four transcription factors of Oct4, Sox2, Nanog and LIN28. According to the method provided by the invention, by independently adding the raw materials and the additives into the separated culture medium, the differentiation after the combination of the pluripotent hematopoietic stem cells and the somatic cells under the conditions of constant temperature and normal oxygen concentration can be simulated and realized; and homing, positioning and self-replication of the pluripotent hematopoietic stem cells are induced by utilizing niche obtained on the surfaces of endoosseous membranes and bone trabecula in a marrow cavity of a mammal or from a commercial mammal blood sinus to form and proliferate the APSC pluripoietic stem cells so as to achieve the purposes of repairing aged and diseased cells and reconstructing functional normal cells and tissues.