39 results about "YAP1" patented technology

Provided herein are methods to increase the culture density and / or thickness of a cellular biomass in a cultivation infrastructure, to improve the culture of cells in the absence of serum in a cultivation infrastructure, and to promote anchorage-independent growth of a cellular biomass in a cultivation infrastructure. The methods comprise inhibiting the HIPPO signaling pathway, for example, by activating YAP1, activating TAZ, and / or inhibiting MOB1, LATS1 kinase, LATS2 kinase, WW45, MST1 kinase, and / or MST2 kinase in the cellular biomass. In some embodiments, the cellular biomass is harvested from the cultivation infrastructure for the formulation of cell-based food products or ingredients, such as animal meat manufactured from cells in an ex vivo process or for therapeutic applications such as organ or tissue transplantation or grafting.

The invention discloses a construction method for a conditional overexpression Yap1 gene mice model at an H11 locus. The construction method comprises the steps: firstly, obtaining an inducible Yap1 overexpression transgenic vector; then, obtaining transgenic F0-generation mice; next, enabling the F0-generation mice to mate with wild-type mice, carrying out passage and line establishment, and thusobtaining F1-generation mice; and carrying out expanding propagation of positive F1-generation mice, then mutually mating, and thus obtaining the conditional overexpression mice Yap1 animal model. The transgenic mice model constructed by the method can be used for studying the function of Yap1 in different tissues and different developmental stages and the action mechanism of Hippo signaling pathways, can also be used for studying the function of the Hippo signaling pathway in animal development, biotumorigenesis, regenerative medicine and other disciplines, and has important guiding significance on biological signal transduction studies.

The invention discloses a YAP1 gene which can be used as a molecular marker for early diagnosis of Alzheimer's disease. The QPCR (quantitative polymerase chain reaction) experiment proves that compared with common people, the YAP1 gene expression in the blood of an Alzheimer's disease patient is obviously enhanced. The RNA (ribonucleic acid) interfering experiment proves that the YAP1 can influence the Abeta mediated neurotoxicity action. According to the research results, a drug capable of inhibiting YAP1 gene expression or inhibiting YAP1 gene expression product functions can be developed, thereby implementing clinical prevention and treatment of Alzheimer's disease.

Binding of the transcriptional co-activator, YAP1, to the transcription factor Oct4, induces Sox2, which is a transcription actor necessary for the self-renewal of stem-like cells from non-small celllung cancer. The WW domain of YAP1 binds to the PPxY motif of Oct4 to induce Sox2. Delivering a peptide corresponding to the WW domain could prevent the induction of Sox2 and stemness. Similarly, peptides and mimetics of the PPxY motif would be able to inhibit stemness. Disclosed are compounds that affect the Yap1:Oct4 interaction.

The invention relates to the technical field of biology, and aims to provide YAP1 gene modified mesenchymal stem cells and a preparation method thereof. The mesenchymal stem cells are primary mesenchymal stem cells modified with overexpressed YAP1 genes, wherein the YAP1 genes are from YAP1 lentiviral vectors or YAP1 plasmid vectors, and the primary mesenchymal stem cells are from any of the following human tissue of placenta, umbilical cords or adipose tissue. The obtained YAP1 gene modified mesenchymal stem cells do not influence the phenotype and the disintegration capacity of MSC; throughthe mesenchymal stem cells modified with overexpressed YAP1 genes, proliferation of the mesenchymal stem cells can be notably promoted, and the yield of cells can be further increased; and therefore,massive mesenchymal stem cells can be quickly obtained for clinical stem cell transplantation treatment.

The invention relates to a gene marker for esophageal cancer prognosis evaluation, and belongs to the technical field of medical molecular biology. The gene marker for esophageal cancer prognosis evaluation is found through screening and analysis of a large number of samples, and comprises the following genes: YAP1, MYC, BAP1, BRIP1, WRN and RB1; and a combined marker for evaluating the total survival time (OS) of esophageal cancer patients is constructed, and can effectively predict high-risk / low-risk esophageal cancer patients.

Aspects of the present disclosure provide compositions and methods for increasing protein production in mammalian cells, e.g. methods of increasing mammalian cell expression of a protein of interest, comprising culturing mammalian cells that overexpress a protein of interest and are modified to overexpress a gene encoding Rab 11 or Yap1, as well as mammalian cells that overexpress a protein of interest and which are modified to overexpress a gene encoding Rab 11 or Yap1.

Immunohistochemical methods and compositions for the typing of molecular subgroups of medulloblastomas are provided. The methods comprise determining a protein expression profile for a sample obtained from a medulloblastoma by detecting expression of GAB 1, filamin A, or at least two biomarker proteins selected from the group consisting of β-catenin, YAP1, GAB1, and filamin A, and typing the medulloblastoma as a WNT pathway tumor, a SHH pathway tumor, or a non-WNT / non-SHH tumor based on this protein expression profile. Kits for typing a medulloblastoma according to these three molecular subgroups are provided. The kits comprise at least two antibodies, wherein each of said antibodies specifically binds to a distinct biomarker protein selected from the group consisting of β-catenin, YAP1, GAB1, and filamin A, and can optionally comprise one or more of instructions for use, reagents for detecting antibody binding to one or more of said biomarker proteins, and one or more positive control samples.

Transgenic oleaginous yeast having increased oil content comprising increased Yap1 transcription factor activity, wherein the increased oil content is compared to the oil content of a non-transgenic oleaginous yeast, are described herein. The increased Yap1 transcription factor activity results from overexpressing a Yap1 transcription factor, by increasing the interaction between the transcription factor and a protein that is capable of activating the transcription factor, or by a combination thereof. Methods of using these yeast strains are also described.

The invention provides a construction method and application of a conditional Yap1 gene knock-in mouse, and relates to the technical field of gene targeting. The nucleotide sequence of a conditional Yap1 knock-in targeting vector is shown as SEQ ID NO.1, and a mouse (Yap1KI) knocked in by the conditional Yap1 knock-in targeting vector can be constructed directly by utilizing the conditional Yap1 knock-in targeting vector. The Yap1 gene knock-in mouse is constructed for the first time, and can be used for constructing a lung cancer model, and an animal model of the Yap1 gene knock-in mouse hasthe characteristic of rapid tumor formation.

The invention belongs to the fields of molecular biology and oncology, and specifically discloses application of tumor-associated antigens RECK protein, RFT2 protein, PTCH1 protein, NOTCH1 protein, FAT4 protein, AJUBA protein, P53 protein and YAP1 protein in preparation of an esophageal cancer detection kit. Especially, the protein is applied to preparation of a kit for early screening esophagealcancer detection, so that the detection efficiency and diagnosis efficiency of clinical esophageal cancer are greatly improved, and the protein is beneficial to wide popularization and application incommon laboratories. When the eight proteins are applied to an ELISA kit, the expression levels of eight TAA antibodies in a serum sample can be detected at the same time, the detection success rate is high, and the technical reproducibility is good. Correspondingly, the molecule capable of specifically detecting the expression level of the combination of RECK, RFT2, PTCH1, NOTCH1, FAT4, AJUBA, P53 and YAP1 can also be used as an esophageal cancer detection test reagent, is applied to an esophageal cancer detection kit, and also has relatively high specificity and sensitivity.

The invention discloses a YAP1 gene and application of an inhibitor in enhancing chemosensitivity of a stem cell of bladder cancer. The YAP1 gene is related to chemosensitivity of the stem cell of bladder cancer. The inhibitor where the YAP1 gene is highly expressed can improve the chemosensitivity of the stem cell of bladder cancer on chemotherapeutics mitomycin C and cis-platinum by down-regulating the expression of the YAP1 gene, so that the inhibiting rate of the chemotherapeutics mitomycin C and cis-platinum on the stem cell of bladder cancer is increased. The inhibitor where the YAP1 gene is highly expressed can be any one or more of siRNA, shRNA, dsRNA, miRNA, cDNA, a small molecular compound, a peptide and an antibody, wherein the small molecular compound is a 2-amido benzoxazole derivative.

Methods for preventing or treating subjects having cancer based on the identification of pharmaceutical agents that target cancer stem cells (CSCs) have been identified. These methods include administering to the subject an effective amount of a COX2 inhibitor and an effective amount of a YAP1 inhibitor. In addition, methods of enhancing chemotherapeutic responses in cancer patients have been discovered and are described herein.

Provided herein is a method for reducing resistance in an individual having a drug resistant cancer, for example, a BRAF inhibitor resistant cancer. A Hippo signaling pathway inhibitor such as a Yes-associated protein 1 (YAP1) inhibitor, a Transcriptional Coactivator with PDZ-binding motif (TAZ) inhibitor, a Transcription enhancer domain (TEAD) inhibitor or a combination of these is administered. Also provided is a method of treating BRAF inhibitor resistance in an individual with a BRAF inhibitor resistant cancer, for example, malignant melanoma, with a Hippo signaling pathway inhibitor, such as Verteporfin, and a BRAF inhibitor.

The invention relates to an application of astrocyte specific METTL3 overexpressed recombinant adeno-associated virus in preparation of drugs for preventing and treating spinal cord injury. The invention successfully constructs the recombinant adeno-associated virus overexpressed by the astrocyte specificity METTL3, and the virus can specifically overexpress the METTL3 protein in the in-vivo astrocyte, modify the m6A level of downstream YAP1 mRNA in a targeted manner, prolong the half-life period, maintain the stability of the mRNA, up-regulate the expression level of the YAP1 protein, promote the activation of reactive astrocyte, and inhibit the expression of the METTL3 protein in the astrocyte. The spinal cord injury function is improved, and a new way is provided for clinical treatment of the spinal cord injury.

The invention discloses a construction method and application of a retina bipolar cytopathy model, and relates to the technical field of medical engineering. Knocking out Yap1 gene, i.e., silencing its expression, in retinal bipolar cells of a target animal can result in the target animal exhibiting characteristics associated with a retinal bipolar cytopathic disease, such as bipolar cell death, primarily manifested by bipolar cell damage, degeneration, progressive thinning of retinal inner nuclear layer, and corresponding pathological changes in retinal outer nuclear layer and other associated cellular layers. Therefore, the target animal knocking out the Yap1 gene can be used as a retinal bipolar cytopathic model, the model can be used for researching pathogenetic process and pathogenesis of the retinal bipolar cytopathic diseases, and provide a basis for in-depth understanding of such diseases.

The invention discloses a phosphorylated antigen polypeptide, a phosphorylated antibody and a preparation method for the phosphorylated antibody, and relates to the technical field of antibody preparation. The phosphorylated antigen polypeptide comprises the phosphorylated antibody which aims at human ERK (Extracellular regulated protein kinases) protein threonine 202 and tyrosine 204, and the amino acid sequence of the phosphorylated antibody is disclosed by SEQ ID NO.1, wherein Thr (threonine) and Try (tyrosine) are mutated into Asp (aspartic acid); and the amino acid sequence of the phosphorylated antibody which aims at YAP1 (Yes-associated protein 1) protein serine 127 is disclosed in SEQ ID NO:2, wherein Ser (serine) is mutated into Asp. A phosphorylated amino acid is mutated into theaspartic acid, the DNA (deoxyribonucleic acid) of a mutant antigen is constructed to a pGEX-6p expression carrier, escherichia coli is used for expressing a GST (glutathione S-transferase) fusion antigen, and after the purified antigen and Freund's adjuvant are mixed, an animal can be directly immunized. The method is simple, has good antigen stability, and is high in solubility and low in cost.

The invention relates to the technical field of drug resistance mechanism research, and discloses a mechanism method of miR-920 targeting YAP1 in esophageal squamous cell carcinoma chemotherapy drug resistance, detection of esophageal squamous cell carcinoma and expression of YAP1 in chemotherapy drug resistance tissues by an immunohistochemical technology, and detection of a chemotherapy drug resistance group by a fluorescent quantitative PCR method. According to the invention, firstly, a dual-luciferase reporter gene experiment is mainly carried out on a cellular level to verify that miR920 can directly regulate expression of YAP1 in a targeting manner; on the basis, a miR920 overexpressed esophageal cancer cell line is constructed by using a lentivirus expression technology, and the characteristics of tumor stem cells can be reduced by miR920 targeted down-regulation of YAP1 through a microsphere culture experiment and other modes. The method of MTS, Westernblot, flow cytometry and the like is further utilized to clarify that the apoptosis resistance of esophageal cancer stem cells can be reversed by miR-920 targeting YAP1. Finally, the effects of the miR-920 and the YAP1 in esophageal cancer chemotherapy drug resistance generation are further determined through detection of clinical specimens. Therefore, an experimental basis and a theoretical basis are provided for screening of the esophageal cancer drug resistance reversal agent.

The invention provides a miRNA marker related to Hu sheep (a kind of sheep mainly raised in Zhejiang Province) muscle cell proliferation and a detection primer and application of the miRNA marker. ThemiRNA marker is miRNA-29a, and the sequence of the miRNA marker is SEQ ID NO. 2. The miRNA targeting YAP1 is predicted through bioinformatics, then a dual luciferase report system, RT-qPCR, CCK-8 andWestern blot technologies are utilized to further verify that the expression of the miRNA-29a plays a significant down-regulation role in the Hu sheep muscle cell proliferation process, and the provided miRNA-29a can be used as a biomarker to be applied to identification of proliferation of Hu sheep muscle cells.

The invention discloses an application of an antisense oligonucleotide ASOYAP1 in inhibition of a plurality of YAP1 positive cancers. A bioinformatics on-line tool is used for analyzing sequence information of a YAP1 protein transcriptome, antisense oligonucleotide sites which have the length of 20 basic groups and can be combined are screened, meanwhile, antisense oligonucleotide is modified, and it is proved that the modified antisense oligonucleotide has the high capacity of inhibiting the level of YAP1 mRNA and protein thereof in cells. The invention provides a new means of nucleic acid drug treatment for clinically treating various cancer cells with high YAP1 expression and important YAP1 effects.

Binding of the transcriptional co-activator, YAP1, to the transcription factor Oct4, induces Sox2, which is a transcription actor necessary for the self-renewal of stem-like cells from non-small celllung cancer. The WW domain of YAP1 binds to the PPxY motif of Oct4 to induce Sox2. Delivering a peptide corresponding to the WW domain could prevent the induction of Sox2 and stemness. Similarly, peptides and mimetics of the PPxY motif would be able to inhibit stemness. Disclosed are compounds that affect the Yap1:Oct4 interaction.

The invention provides a building method of spontaneous squamous cell lung carcinoma mouse models, and relates to the technical field of building of mouse models. According to the building method of the spontaneous squamous cell lung carcinoma mouse models, by designing targeting vectors with conditional Yap1 knocked in, mice (Yap1KI) with the conditional Yap1 knocked in are built, due to tumor-promoting action of Yap1, a spontaneous squamous cell lung carcinoma has the character of fast tumor formation, then according to standard design of transgenic mice, negative selection markers (DTA) andpositive selection markers (Neo) are also introduced into the targeting vectors, thus positive clones can be conveniently and fast obtained, and by conducting hybridization on mice with Trp53 genes knocked out and the Yap1KI mice, Yap1KI / Trp53KO mice are obtained at last, so that building of the spontaneous squamous cell lung carcinoma mouse models is completed.