33 results about "DLK1" patented technology

The present invention relates to antisense oligonucleotides that modulate the expression of and / or function of Delta-like (1) homolog (DLK1), in particular, by targeting natural antisense polynucleotides of Delta-like (1) homolog (DLK1). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of (DLK1).

The invention relates to key genes, microRNAs and other non-coding RNAs, or a combination thereof used for identifying or regulating cell pluripotency. The invention is characterized in that the key genes, microRNAs, other non-coding RNAs or a combination is highly expressed in the stem cell with complete pluripotency and the expression is obviously repressed or silent in the stem cell without complete pluripotency. The genes, microRNAs and other non-coding RNAs are the genes, microRNAs and other non-coding RNAs positioned in the chromosome imprinting region named as Dlk1-Dio3 on the long arm of mouse chromosome 12 and the genes, microRNAs and other non-coding RNAs in the genome collinearity regions of other mammals, which have 70%-100% of homology. The invention also relates to applications of the genes, microRNAs and other non-coding RNAs, or the combination thereof used for identifying the pluripotency of the stem cell and regulating cell pluripotency, applications in stem cell typing, applications for regulating the cell pluripotency and the pluripotency state and level of the cell, applications in disease treatment, and applications in the drug target development of tumor treatment or the development of antitumor drugs.

The present invention discloses an application of DLK1 gene for preparing a RNA interference medicine for remedying the primary liver cancer and provides a siRNA sequence which considers the DLK1 gene as a target point. The present invention comprises the carrier of the siRNA sequence which considers the DLK1 gene as the target point and a host cell and also provides a biological chip and a kit for remedying and diagnosing the liver cancer. The DLK1 is considered as the target point for preparing the medicine for remedying the primary liver cancer, which ensures that the liver cancer is likely to be conquered.

The invention relates to application of a DLK1 (delta drosophila homolog-like1) gene in preparation of a gastrointestinal stromal tumor diagnostic reagent. The new discovery of the invention can be applied to development of drugs or kits based on RT-PCR, PCR, immunodetection, in-situ hybridization, gene chip and other technologies for diagnosis of gastrointestinal stromal tumors. The invention puts forward for the first time that detection based on DLK1 gene and its products in serum / tissue can be utilized to achieve early diagnosis of GIST or carry out prognosis on GIST patients undergoing surgical treatment, the high-throughput chip screening + real-time quantitative PCR validation established on the gene level and large sample clinical sample validation on the protein level have reliable result, and DLK1 can significantly distinguish tumors and nontumors, high-risk tumors and low-risk tumors both on the gene level and protein level, and the result repeatability is good. Thus, DLK1 gene has good clinical application value.

The present invention relates to the use of antibodies recognizing Fetal Antigen-1 (FA1 / dlk1) for the detection and isolation of cell populations in mammalian muscle. In one embodiment, myogenic progenitor cells are detected in developing, diseased or regenerating muscle. In another embodiment, muscle stem and progenitor myogenic progenitor cells are isolated from muscle tissue or from cultures containing muscle cells. The isolated cells may be used for transplantation, drug screening, production of cell type specific antibodies, and gene therapy and discovery. Transplantation of these cells may provide treatments for degenerative diseases of muscle, and for regeneration of muscle following trauma or ischemia such as myocardial infarction.

This application relates to a method for selecting an induced pluripotent stem cell (iPS), the method comprising: selecting an iPS cell that expresses a gene in the Dlk1-Dio3 cluster from a population of iPS cells. The method further comprises: comparing the gene expression profile determined for an iPS cell with the gene expression profile determined for an embryonic stem cell; identifying a gene that is differentially expressed in the embryonic stem cell as compared to the iPS cell; and selecting the desired iPS cell from a population of iPS cells.

Methods and tools are provided for detecting and predicting lung cancer. The methods and tools are based on epigenetic modification due to methylation of genes in lung cancer or pre-lung cancer. The tools can be assembled into kits or can be used seperately. Genes found to be epigentically silenced in association with lung cancer include ACSL6, ALS2CL, APC2, ART-S1, BEX1, BMP7, BNIP3, CBR3, CD248, CD44, CHD5, DLK1, DPYSL4, DSC2, EDNRB, EPB41L3, EPHB6, ERBB3, FBLN2, FBN2, FOXL2, GNAS, GSTP1, HS3ST2, HPN, IGFBP7, IRF7, JAM3, LOX, LY6D, LY6K, MACF1, MCAM, NCBP1, NEFH, NID2, PCDHB15, PCDHGA12, PFKP, PGRMC1, PHACTR3, PHKA2, POMC, PRKCA, PSEN1, RASSF1A, RASSF2, RBP1, RRAD, SFRP1, SGK, SOD3, SOX17, SULF2, TIMP3, TJP2, TRPV2, UCHL1, WDR69, ZFP42, ZNF442, and ZNF655.

The invention discloses ribonucleic acid taking a liver cancer related gene DLK1 as a target spot, which has the following oligonucleotide sequences: siRNA:S:GAGCUACGAGUGUCUGUGCAAGCdTdT (SEQ IDNO:10); AS:GCUUGCACAGACACUCGUAGCUCdTdT (SEQ ID NO:11). In addition, the invention also discloses application of the ribonucleic acid taking the liver cancer related gene DLK1 as the target spot in preparing an RNA interference medicament for curing primary liver cancer.

The invention relates to a kit for detecting hybridization in situ for DLK1 gene of early cancer, which comprises a hybridization probe and a marker, wherein the sequence of the hybridization probe is expressed as SEQ ID NO.1. The invention also provides a method for detecting hybridization in situ for the DLK1 gene. The detection kit and the detection method can detect expression amount of the DLK1 gene in early canceration cells or in canceration cells at the level of genes, can detect change information of the DLK1 gene before generation of protein markers and more before oncogenesis, and really perform early diagnosis of tumor. The kit has the advantages of high sensitivity and strong specificity. The detection method has the advantage of convenient and simple operation, and can be generally used and popularized in hospitals above districts.

The invention discloses a method for extracting active components of skin stem cells. The method comprises the steps of adding human skin tissues to a normal saline mixed digestive fluid of a type I collagenase and a type II collagenase added with a PMSF protease inhibitor for constant temperature shaking table culture; stopping digestion by a culture medium M199 containing 10% by volume of fetal calf serum; performing centrifuging and precipitate collection; performing cell re-suspension of normal saline to obtain compound skin stem cells; and screening out Dlk1<+> / Scal<-> skin stem cells by applying magnetic bead marked Dlk antibody and Scal antibody. The purity of the Dlk1<+> / Scal<-> skin stem cells extracted with the method can reach 95.8%; and the skin stem cells are large in quantity, high in cell survival rate and high in activity, are applied to drugs for treating hair loss, and have a remarkable hair growth effect.

The present invention relates to a pharmaceutical composition for preventing or treating a muscular disease or cachexia, comprising, as an active ingredient, a miRNA located in Dlk1-Dio3 cluster or avariant thereof. In the present invention, it has been found that expression of miRNAs located in the Dlk1-Dio3 cluster is decreased as age increases. In particular, in a case where most of the miRNAsare over-expressed in fully differentiated myotubes, it has been confirmed that the diameter of the myotubes increases. In addition, also in a tumor-induced cachexia mouse model, it has been confirmed that cachexia was improved by inhibiting Atrogin-1 protein. Accordingly, the miRNA located in the Dlk1-Dio3 cluster or a variant thereof can be usefully utilized for the treatment and prevention ofan Atrogin-1 - dependent muscular disease and cachexia.

The present invention relates to an antibody against delta-like 1 homolog (Drosophila) (DLK1) or an antigen-binding fragment thereof, a nucleic acid encoding the same, a vector comprising the nucleic acid, a cell transformed with the vector, a method for producing the antibody or an antigen-binding fragment thereof, an antibody drug conjugate (ADC) comprising the same, a pharmaceutical composition for treating cancer, a composition for diagnosing cancer, and a chimeric antigen receptor (CAR) and a T-cell engager comprising the same.

The invention discloses a CAR-T construction method with antigens DLK1 related to liver cancers as targets. According to the method, single-chain antibodies of the antigens DLK1 related to the liver cancers are selected to be recombined into T cells, the genetically-engineered T cells are further activated, and therefore the lethality of the T cells on liver cancer cells is improved; according tothe single-chain antibodies of DLK1, RNA is obtained from a hybridoma cell line which secretes monoclonal antibodies of DLK1, and then cDNA is obtained after reverse transcription; PCR primers for a heavy-chain variable area and a light-chain variable area are designed; with cDNA as a template, the single-chain antibodies of DLK1 are obtained by amplifying and splicing the heavy-chain and light-chain variable areas. A detection method has relative sensitivity and specificity, and the side effects of recognizing normal cells by the T cells can be reduced. The CAR-T construction method providesa certain research basis for a CAR-T cell immunotherapy in the aspect of the immunotherapy of the liver cancers.

The present invention relates to: novel antibody-drug conjugates (ADC) targeting DLK1; active metabolites of the ADCs; a method for producing the ADCs; a use of the ADCs for treating and / or preventingdiseases; and a use of the ADCs for producing medicaments for treating and / or preventing diseases, more specifically, hyperproliferative and / or angiogenic diseases such as cancer. More particularly,the present invention relates to an antibody-drug conjugate comprising an antibody binding to DLK1 or an antigen-binding fragment thereof, and to a pharmaceutical composition including same.

The invention relates to key genes, microRNAs and other non-coding RNAs, or a combination thereof used for identifying or regulating cell pluripotency. The invention is characterized in that the key genes, microRNAs, other non-coding RNAs or a combination is highly expressed in the stem cell with complete pluripotency and the expression is obviously repressed or silent in the stem cell without complete pluripotency. The genes, microRNAs and other non-coding RNAs are the genes, microRNAs and other non-coding RNAs positioned in the chromosome imprinting region named as Dlk1-Dio3 on the long arm of mouse chromosome 12 and the genes, microRNAs and other non-coding RNAs in the genome collinearity regions of other mammals, which have 70%-100% of homology. The invention also relates to applications of the genes, microRNAs and other non-coding RNAs, or the combination thereof used for identifying the pluripotency of the stem cell and regulating cell pluripotency, applications in stem cell typing, applications for regulating the cell pluripotency and the pluripotency state and level of the cell, applications in disease treatment, and applications in the drug target development of tumor treatment or the development of antitumor drugs.

Methods and tools are provided for detecting and predicting lung cancer. The methods and tools are based on epigenetic modification due to methylation of genes in lung cancer or pre-lung cancer. The tools can be assembled into kits or can be used separately. Genes found to be epigenetically silenced in association with lung cancer include ACSL6, ALS2CL, APC2, ARTS-1, BEX1, BMP7, BNIP3, CBR3, CD248, CD44, CHD5, DLK1, DPYSL4, DSC2, EDNRB, EPB41L3, EPHB6, ERBB3, FBLN2, FBN2, FOXL2, GNAS, GSTP1, HS3ST2, HPN, IGFBP7, IRF7, JAM3, LOX, LY6D, LY6K, MACF1, MCAM, NCBP1, NEFH, NID2, PCDHB15, PCDHGA12, PFKP, PGRMC1, PHACTR3, PHKA2, POMC, PRKCA, PSEN1, RASSF1A, RASSF2, RBP1, RRAD, SFRP1, SGK, SOD3, SOX17, SULF2, TIMP3, TJP2, TRPV2, UCHL1, WDR69, ZFP42, ZNF442, and ZNF655.

The invention discloses ribonucleic acid taking a liver cancer related gene DLK1 as a target spot, which has the following oligonucleotide sequences: siRNA:S:GAACCUGCUGCUUCAGUACAAdTdT (SEQ IDNO:20); AS:UUGUACUGAAGCAGCAGGUUCdTdT (SEQ ID NO:21). In addition, the invention also discloses application of the ribonucleic acid taking the liver cancer related gene DLK1 as the target spot in preparing an RNA interference medicament for curing primary liver cancer.

The invention discloses an ELISA kit for detecting human serum DLK1 protein and its application. The antigen of the ELISA kit of the present invention adopts recombinant human DLK1 antigen. The detection method of the present invention has high sensitivity and specificity, and can quickly detect the expression of DLK1 and liver cancer and hepatoblastoma. The ELISA kit of the present invention has broad application in the prevention and diagnosis of liver cancer and hepatoblastoma Application prospects.

The invention discloses a method for detecting the fat coverage rate of Simmental cattle carcass by DLK1 gene markers. Using the DNA sequence of the DLK1 gene of Chinese Simmental cattle as a template, a pair of primers are designed, and a strict PCR reaction system is carried out to carry out PCR of samples Amplification and electrophoresis, screening by sequencing and finding 2 SNP sites; the restriction endonuclease for screening DLK1-478 C / T sites in the population is Msp I, and the cleavage base limited by this enzyme is CCGG; screening The restriction endonuclease of DLK1‑609 T / G site is Ase I; the individuals carrying DLK1‑478 C / T site T allele and TT mutation genotype in Chinese Simmental cattle population have higher fat coverage and better marbling; and individuals with the G allele and GG mutant genotype carrying the DLK1‑609 T / G locus had higher fat coverage. The two mutation sites of the DLK1 gene of the present invention can be used as genetic markers predicting the fat coverage rate and marbling traits of Chinese Simmental cattle, and used for early marker-assisted selection of beef cattle.

The invention discloses sgRNA for up-regulating non-coding RNA expression in a human DLK1-DIO3 imprinted domain, a recombinant plasmid and a cell line. The sgRNA sequence is shown as SEQ ID NO. 1, andspecifically as 5'-TTTATATGGAGGCGCAGAAG-3'; the method comprises the steps of: synthesizing a nucleic acid fragment of a sgRNA sequence and inserting into a multiple cloning site of the MS2-P65-HSF1expression plasmid vector and transforming, and selecting a monoclonal strain, extracting the recombinant plasmid of MS2-P65-HSF1-sgRNA-DLK1-DIO3, and transfecting the recombinant plasmid to the target cell line which is pre-transfected with dCAS9-VP64 plasmid to obtain the cell strain for up-regulating non-coding RNA expression in the DLK1-DIO3 imprinted domain. The application can rapidly, easily and accurately up-regulate the expression of non-coding RNA on the DLK1-DIO3 imprinted domain of the cell strain.

The invention discloses ribonucleic acid taking a liver cancer related gene DLK1 as a target spot, which has the following oligonucleotide sequences: siRNA:S:CAGGCAAUUUCUGCGAGAUCGUGdTdT (SEQ IDNO:6); AS:CACGAUCUCGCAGAAAUUGCCUGdTdT (SEQ ID NO:7). In addition, the invention also discloses application of the ribonucleic acid taking the liver cancer related gene DLK1 as the target spot in preparing an RNA interference medicament for curing primary liver cancer.

The present invention relates to a pharmaceutical composition for preventing or treating a muscular disease or cachexia, comprising, as an active ingredient, a miRNA located in Dlk1-Dio3 cluster or a variant thereof. In the present invention, it has been found that expression of miRNAs located in the Dlk1-Dio3 cluster is decreased as age increases. In particular, in a case where most of the miRNAs are over-expressed in fully differentiated myotubes, it has been confirmed that the diameter of the myotubes increases. In addition, also in a tumor-induced cachexia mouse model, it has been confirmed that cachexia was improved by inhibiting Atrogin-1 protein. Accordingly, the miRNA located in the Dlk1-Dio3 cluster or a variant thereof can be usefully utilized for the treatment and prevention of an Atrogin-1-dependent muscular disease and cachexia.

The present invention discloses human gene Delta like 1 homolog (DLK1) and the application of its coded protein in diagnosing and treating liver cancer. The human DLK1 gene located in 14q32 of chromosome encodes one protein containing 383 amino acids. The human DLK1 gene has expression in liver cancer more obviously than in nearby tissues. Researching on the liver cancer relative human LLK1 gene and its coded protein in depth may find out new mechanism of liver cancer. At the same time, they may become the new target site for liver cancer treatment and may provide important reference for developing liver cancer treating medicine.

The invention provides application of a urine protein delta homolog 1 (DLK1) and a polypeptide fragment thereof, and particularly relates to application of the urine protein delta homolog 1 and the polypeptide fragment thereof in preparation of preparations for diagnosis and treatment of gestational diabetes patients, monitoring and evaluation of blood sugar and metabolic states, mechanism research and the like. Researches prove that the urine protein delta homolog 1 and the polypeptide fragment thereof are reduced along with the increase of the HbA1c level in gestational diabetes patients, have statistical significance in difference between a BM1 group with the HbA1c being less than or equal to 5.1% and a BM2 group with the HbA1c being greater than 5.1%, and can be used for diagnosis and treatment of gestational diabetes patients, monitoring and evaluation of blood sugar and metabolic states, mechanism research and other application detection for various purposes. The advantages of noninvasive acquisition, large-scale repeated sampling and convenient preservation of a urine sample are exerted, and the protein delta homolog 1 and the polypeptide fragment thereof are detected by utilizing the urine sample.

The invention discloses ribonucleic acid taking a liver cancer related gene DLK1 as a target spot, which has the following oligonucleotide sequences: siRNA:S:CAGCCGGCUUCAUCGACAAGAdTdT (SEQID NO:8); AS:UCUUG UCGAUGAAGCCGGCUGdTdT (SEQ ID NO:9). In addition, the invention also discloses application of the ribonucleic acid taking the liver cancer related gene DLK1 as the target spot in preparing an RNA interference medicament for curing primary liver cancer.

The invention discloses a CAR-T construction method targeting the liver cancer-associated antigen DLK1. In the present invention, the single-chain antibody of liver cancer-associated antigen DLK1 is recombined into T cells, and the T cells after genetic engineering are further activated, thereby enhancing the lethality of T cells to liver cancer cells; the single-chain antibody of DLK1 is secreted Obtain RNA from the hybridoma cell line of DLK1 monoclonal antibody, and obtain cDNA after reverse transcription; design PCR primers for the variable region of the heavy chain and the variable region of the light chain; obtained by amplification and splicing of chain variable regions. The detection method of the invention has relative sensitivity and specificity, and can reduce the side effects caused by T cells recognizing normal cells. The present invention provides a certain research basis for CAR-T cell immunotherapy in liver cancer immunotherapy.

The invention relates to application of a DLK1 / MEG3 locus in detection and treatment of growth hormone adenoma, and quick and accurate detection of the growth hormone adenoma can be realized by identifying the DLK1 / MEG3 locus and / or an expression product of the DLK1 / MEG3 locus.