77 results about "Transcriptional repression" patented technology

The present invention provides RNA-guided endonucleases, which are engineered for expression in eukaryotic cells or embryos, and methods of using the RNA-guided endonuclease for targeted genome modification in in eukaryotic cells or embryos. Also provided are fusion proteins, wherein each fusion protein comprises a CRISPR / Cas-like protein or fragment thereof and an effector domain. The effector domain can be a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Also provided are methods for using the fusion proteins to modify a chromosomal sequence or regulate expression of a chromosomal sequence.

According to the present invention, a gene having a novel function that can cause an increase or decrease in seed protein content is searched for. A chimeric protein obtained by fusing a transcription factor consisting of a protein comprising an amino acid sequence shown in any of the even-numbered SEQ ID NOS: 1 to 76 and a functional peptide capable of converting an arbitrary transcription factor into a transcriptional repressor or a transcription factor consisting of a protein comprising an amino acid sequence shown in any of the even-numbered SEQ ID NOS: 77 to 84 is expressed in a plant.

Parkinson's disease is caused by the preferential loss of substantia nigra dopamine neurons. A Parkin Interacting Substrate, PARIS (ZNF746) is identified. The levels of PARIS are regulated by the ubiquitin proteasome system via binding to and ubiquitination by the E3 ubiquitin ligase, parkin. PARIS is a KRAB and zinc finger protein that accumulates in models of parkin inactivation and in human brain Parkinson's disease patients. PARIS represses the expression of the transcriptional co-activator, PGC-1α and the PGC-1α target gene, NRF-1 by binding to insulin response sequences in the PGC-1α promoter. Conditional knockout of parkin in adult animals leads to progressive loss of dopamine (DA) neurons that is PARIS dependent. Overexpression of PARIS causes selective loss of DA neurons in the substantia nigra, which is reversed by either parkin or PGC-1α co-expression. The identification of PARIS provides a molecular mechanism for neurodegeneration due to parkin inactivation.

The invention relate to methods, compositions, and kits for reprogramming a cell. In one embodiment, the invention relates to a method for inducing the expression of at least one gene that contributes to a cell being pluripotent or multipotent. In yet another embodiment, the method comprises inhibiting the expression of a gene that codes for a protein involved in transcriptional repression. In yet another embodiment, the invention relates to a reprogrammed cell or an enriched population of reprogrammed cells that can have characteristics of an ES-like cell, which can be re- or trans-differentiated into a differentiated cell type.

A gene having novel functions is searched for, by which plant weight (that is, biomass level) can be increased and by which substance productivity can be increased or decreased. A chimeric protein is expressed in which a transcriptional factor comprising the amino acid sequence shown in SEQ ID NO: 2, 4, or 6 is fused to a functional peptide that converts an arbitrary transcriptional factor into a transcriptional repression factor.

The invention discloses a bacillus subtilis polygene editing and expression adjusting and controlling system based on CRISPR Cpf1. Through a constructed Cpf1 expression vector pHT-XCR6 and a constructed crRNA array expression vector pcrF11, complete knockout of 2 genes, basic group modification of 6 genes and knock-in of 1 gene can be completed at one time. The vector pHT-XCR6 contains NgAgo protein, and is used for promoting recA mediated homologous recombination. Besides, a vector pLCg6-dCpf1-remA (used for integral expression of a Cpf1 mutant dCpf1 inactivated by deoxyribonuclease and fusion protein of an activating transcription factor remA on a bacillus subtilis genome) and a vector pcra3 (used for integral expression of a crRNA array on the bacillus subtilis genome) are constructed,and can be used for performing transcription inhibition and activation on different genes. The invention further establishes an economic efficient crRNA array assembling method of which the name is SOMACA (Synthetic Oligos Mediated Assembly of crRNA Array), and through synthesizing short-single-strand DNA (60nt ), a needed crRNA array is inserted in the vector pcrF11 or pcra3 and is used for guiding Cpf1 or dCpf1-remA for gene editing and expression adjustment and control.

The invention aims at developing a novel DNA (deoxyribonucleic acid) inserting reagent with good antitumor activity. A cyclometalated single-core ruthenium (II) complex is synthetized. The complex is stable in structure; the water-solubility is better than that of a common organic small molecular reagent at present; good DNA transcription inhibitory activity is represented. A cytotoxicity test indicates that the cyclometalated ruthenium (II) complex has significant growth inhibition effects on cancer cells of 11 different tissue parts of a human body; the inhibitory activity is much better than that of cis-platinum.

The invention relates to the technical field of plant gene engineering, in particular to a method for increasing the grain number per ear and reducing plant height by use of rice SNB genes. The nucleotide sequence of SNB genes is shown as SEQ ID NO:1. Artificially mutant rSNB genes are obtained by changing the binding site of microRNA172 in the SNB genes, the nucleotide sequence of the rSNB genes is shown as SEQ ID NO:2, and the sequence of protein encoded by the rSNB genes is shown as SEQ ID NO:3. The grain number per ear of rice is remarkably increased and the plant height of the rice is notably reduced through overexpression of rSNB, and further studies prove that SNB encoded protein has the activity of a transcription suppressor. The functions of the SNB genes on increasing the grain number per ear of the rice and reducing the plant height of the rice are disclosed, and a novel rice genetic improvement method is also provided.

This invention relates to a thirteen amino acid polypeptide sequence which is able to autonomously function as a transcription repression domain through its ability to bind mSin3A. Disclosed herein are compounds and methods for regulating transcription of a selected gene. Compounds include fusions of this repression domain to a DNA-binding domain, such as a polyamide or zinc finger domain. Methods are provided for constructing and for using such compounds to regulate transcription of a selected gene.

This invention relates to inhibition of hepatitis gene expression. More specifically, the invention relates to a method of using RNA sequences to inhibit Hepatitis B and C Virus replication. Expression cassettes that include DNA sequences derived from endogenous micro RNAs (miRs) are used in the method and are transcribed by Pol Il promoters, and then processed to generate sequences that are specific to target hepatitis virus sequences (RNAi effecter sequences). The RNAi effecter sequences can target the selected hepatitis virus sequences resulting in gene silencing or transcriptional inhibition of the hepatitis virus gene. The expression cassettes may be delivered in vitro or in vivo to host cells. A pharmaceutical composition containing the expression cassettes is also claimed.

The invention provides pCasSA plasmid and an application thereof. The sequence of the pCasSA plasmid is as shown in SEQ ID NO: 1. The pCasSA plasmid is capable of (1) effectively and rapidly editing genomes of various staphylococcus aureus strains, including gene knockout, single base mutation and gene insertion, and (2) having an efficient transcription inhibition function on target genes. The technique has wide application prospects in such aspects as staphylococcus aureus infection treatment, medicine target discovery, medicine development and staphylococcus aureus physiological study.

The present invention relates to the identification of the genomic promoter region of the human and mouse telomerase RNA gene. Telomerase activity is necessary for the unrestricted proliferative capacity of many human cancers. It is proposed that mutation or dysregulation of the telomerase repression pathway may cause reactivation or upregulation of telomerase expression in cancer. The invention provides details of elements important for the regulation of telomerase RNA genes, including the Sp family of transcription factors. There is further provided methods for screening elements having the ability for suppressing telomerase RNA gene promoter activity and use of such elements in the treatment of cancers. In addition, evidence is also provided for the development of new transcription based therapies for cancer and for genetic approaches to targeting therapeutic genes to cancer cells. Namely, (1) transcriptional repression and the disruption of signal transduction pathways regulating telomerase activation. (2) Tumour specific gene expression for genetic therapy via telomerase RNA gene promoters.

The invention discloses an aptazyme modified sgRNA carrier with theophylline regulated expression. Aptazyme P1-F5 is inserted to tetraloop and stem loop 2 positions of an expressed sgRNA-AZ 2.0 skeleton; by means of Kpn I and EcoR I sites, and EcoR I and Spe I sites, a U6 promoter and the sgRNA-AZ 2.0 skeleton are connected into the carrier pUC19 / EKSHL in order to obtain the carrier pU6-sgRNA-AZ 2.0; using Bsa I for enzyme digestion of the carrier pU6-sgRNA-AZ 2.0, and using cohesive end design and connecting an sgRNA sequence directing at the target gene so as to obtain the pU6-sgRNA-AZ 2.0-target site. The carrier can effectively mediate the genome editing function of Cas9 protein, and bring the editing activity under regulation of theophylline, also can mediate the transcription inhibition of dCas9-KRAB protein to the target gene, and enables regulation of inhibitory activity by theophylline.

The present invention provides methods for discovering agents that are effective in reversing epigenetic silencing by inhibiting the interaction of methyl-binding (MBD) proteins with methylated genomic DNA. Also provided are methods for reactivating silenced genes having CpG island hypermethylation along with methods for treatment and prevention of diseases, such as cancer and sickle cell anemia, by administering an agent that modulates methyl-binding domain (MBD) protein-mediated transcriptional repression, thereby increasing gene transcription to prevent or treat disease. Additionally, compounds identified by the present invention useful for treatment and prevention of diseases, such as cancer and sickle cell anemia, are provided.

The invention relates to fusion proteins capable of acting as transcriptional repressors of cytokinin signaling, to polynucleotides encoding these fusion proteins, to vectors and cells comprising these polynucleotides, and to transgenic plants and parts thereof comprising these polynucleotides, vectors, and cells.The invention further relates to a process for making these transgenic plants and to the use of these transgenic plants for producing seeds of enhanced size, with enhanced seed filling, with reduced seed loss and / or with more rapid germination, and / or for producing a live root system with increased root mass, root length and / or root branching. The invention also relates to a method for enhancing the seed size, for enhancing seed filling, for reducing seed loss, and / or for reducing germination time and / or reproduction time, and / or for enhancing the root mass, root length and / or root branching of a plant and to seeds obtainable by the methods of the present invention.

The invention relates to genetically modified plants with an altered seed phenotype, in particular increased seed size. The invention relates to a plant that does not produce a functional NGAL2 polypeptide or functional NGAL2 and NGAL3 polypeptides. NGAL2 and NGAL3 are members of the RAV family and comprise a B3 DNA- binding domain and a transcriptional repression motif.

The invention relate to methods, compositions, and kits for reprogramming a cell. In one embodiment, the invention relates to a method for inducing the expression of at least one gene that contributes to a cell being pluripotent or multipotent. In yet another embodiment, the method comprises inhibiting the expression of a gene that codes for a protein involved in transcriptional repression. In yet another embodiment, the invention relates to a reprogrammed cell or an enriched population of reprogrammed cells that can have characteristics of an ES-like cell, which can be re- or trans-differentiated into a differentiated cell type.

The invention discloses a gene expression regulation and control method and system based on Type I-F CRISPR / Cas. The first study shows that a Cascade compound, such as a PaeCascade compound, belonging to a Type I-F CRISPR / Cas system can be efficiently combined with a target site in a mammalian cell; and meanwhile, the Type I-F CRISPR / Cas system is subjected to mammalian expression system optimization, so that a transcription activation factor or a transcription inhibition factor can be efficiently recruited to the target site in the mammalian cell, and a transcription activation system anda transcription inhibition system are successfully constructed. The application of the Type I-F CRISPR / Cas system in the aspect of gene regulation and control of the mammalian cell becomes possible,and a necessary tool is provided for gene transcription regulation and control based on the Type I-F CRISPR / Cas system.

The invention relates to the field of biotechnology and discloses a high-proliferation specific recombination eucaryotic cell promoter PCNATAI; an eucaryotic cell operating system PCNATAI-LoxP-stop-LoxP taking a PCNATAI as a core transcription initiator and a LoxP-stop-LoxP as a transcriptional repression element; and a set of general scheme for eliminating the repression effect of the LoxP-stop-LoxP to the transcription process of the PCNATAI promoter through a Cre recombinase provided by a tumor cell specific dependent model trans form and enabling the downstream exogenous tumor suppressor gene or the suicide gene to be effectively and specifically expressed in the malignant tumor cell in a tumor cell specific pattern and a proliferation specific pattern. The scheme not only can remarkably improve the tumor inhibition rate during the gene therapy of the malignant tumor or the expression efficiency of the suicide gene in the malignant tumor cell, but also can ensure the security of the gene therapy.

The invention provides a PLK1 inhibitor, and a preparation method and an application thereof. The PLK1 inhibitor is a compound represented by general formula (I) shown in the description, and stereoisomers or pharmaceutically acceptable salts thereof. All groups in the formula (I) are defined as in the description. The PLK1 inhibitor can specifically bind to a DNA sequence in a PLK1 gene transcription promoter region, represented by SEQ: NO:1, in a high strength manner, inhibits transcription of the PLK1 gene, inhibits the expression of a PLK1 protein, causes tumor cell growth inhibition or apoptosis, can traverse through a cell membrane and a nuclear membrane and resist nuclease hydrolysis, and also solves the drug resistance problem of micro-molecular kinases inhibitors.

The invention discloses a method for detecting the inhibiting ability of silent GPC-3 (glypican-3) gene transcription on a hepatocellular carcinoma transplanted tumor in a nude mouse. The method includes: 1) designing and synthesizing 4 pairs of miRNA oligomeric single-strand DNA specific to a target gene consensus sequence, then synthesizing the corresponding dsDNA, inserting the dsDNA into a vector to construct 4 recombinant plasmids, and picking out the recombinant plasmid with the highest interference efficiency through fluorescence quantitative PCR; 2) transfecting the recombinant plasmid with the highest interference efficiency into HepG2 cells to construct transfected HepG2 cells, and performing screening, thus obtaining a stably transfected HepG2 cell; 3) inoculating the stably transfected HepG2 cell to the nude mouse subcutaneously to construct a human hepatocellular carcinoma tumor-bearing nude mouse model; and 4) then determining the growth state of the stably transfected HepG2 cell in the human hepatocellular carcinoma tumor-bearing nude mouse model. The method provided by the invention can be employed to detect the inhibiting ability of silent GPC-3 gene transcription on a hepatocellular carcinoma transplanted tumor in the nude mouse, and can be used for research of GPC-3 on transplanted tumors.

Method of increasing protein content in a eukaryotic cell comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site; method of producing a plant with increased proteincontent comprising crossing and selecting for increased protein content; method of increasing resistance to a pathogen or a pest in a plant comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site, alone or in further combination with expressing QQS in the plant; method for producing a plant with increased resistance to a pathogen or a pest comprising crossing andselecting for increased resistance to the pathogen or the pest; a cell, collection of cells, tissue, organ, or organism in which the NF-YC4 gene comprises a promoter comprising a transcriptional repressor binding site that has been modified so that the transcriptional repressor cannot prevent transcription of the NF-YC4; hybrid plants; and seeds.

This invention provides methods of screening for compounds that inhibit herpesviral transcription and replication. The methods comprise screening test compounds for ability to enhance the activity of homeodomain transcription factor PDX1 in repressing transcription of herpesviral genes (e.g., the IE gene of cytomegalovirus). Transcriptional repression by PDX1 can be monitored using an expression vector comprising a reporter gene operably linked to a PDX1-binding, upstream transcription regulatory sequence of the herpesvirus. The invention further provides methods and pharmaceutical compositions for stimulating PDX1-mediated transcriptional repression in a subject and for treating diseases and conditions associated with herpesviral infection.

The invention discloses a conditionally replicating adenovirus vector for virus replication regulated by a TetR-KRAB-mediated transcription inhibition type Tet-On system, adenovirus E1 region comprises the following components in the following connection sequence: 5'-CMV promoter, TetR-KRAB gene, promoter TRE3G-E1b Pro containing a tetracycline responsive element, EcoR I enzyme cutting site, E1B Delta 55KD protein-deleted adenovirus E1A-E1B19KD gene sequence, enzyme cutting site Spe I and mRNA transcription termination signal SV40polyA-3'; the CMV promoter expresses transcription inhibitor TetR-KRAB; a gene sequence inserted into the two enzyme cutting sites of the EcoR I and the Spe I is a gene fragment from adenovirus E1A translation initiation site to adenovirus E1B19KD protein termination codon, the promoter TRE3G-E1b Pro containing the tetracycline responsive element expresses adenovirus E1A gene, adenovirus E1B19KD gene is expressed by the own promoter of the adenovirus E1B19KD, and the packed adenovirus is named as Ad5-CMV-TetR-KRAB-TRE3G-E1bPro-Delta 55KD. Tests show that the virus can be loaded into mesenchymal stem cells for application in study of tumor therapy.

The invention relate to methods, compositions, and kits for reprogramming a cell. In one embodiment, the invention relates to a method comprising inducing the expression of at least one gene that contributes to a cell being pluripotent or multipotent. In yet another embodiment, the method comprises delivering a transcription factor to a cell and exposing said cell to an agent that inhibits the activity, expression, or activity and expression of a gene, which codes for a protein, or a protein involved in transcriptional repression, and selecting a cell, wherein differentiation potential has been restored to said cell. In yet another embodiment, the invention relates to a reprogrammed cell and an enriched population of reprogrammed cells that can have characteristics of an ES-like cell can be re- or trans-differentiated into various differentiated cell types.

The present invention relates to a recombinant adenovirus capable of regulating angiogenesis, which comprises (a) an adenoviral UR (inverted terminal repeat) nucleotide sequence; and (b) a transcription regulatory sequence for a VEGF-A (vascular endothelial growth factor-A) gene comprising (i) a nucleotide sequence encoding a DNA binding domain comprising a zinc finger domain to bind to a site in a VEGF-A promoter sequence as set forth in nucleotides 1-2362 of SEQ ID NO:1, and (ii) a transcription activation domain or a transcription inhibitory domain linked to the nucleotide sequence encoding the DNA binding domain; and a pharmaceutical composition comprising the recombinant adenovirus.

The invention discloses a transcriptional repression domain as well as encoding genes and application thereof in regulating stress tolerance of plants. The transcriptional repression domain comprisesamino acid sequences as shown in SEQ ID NO:1 or SEQ ID NO:2 or SEQ ID NO:3. Since the transcriptional inhibitory activity of the existing SRDX is too strong, transgenic plants in which certain transcription factors are fused with the SRDX may not survive. By contrast, inhibitory sequences provided by the invention have transcriptional inhibitory activity, but the transcriptional inhibitory activity is lower than that of the SRDX; and the transcriptional repression domain is of great scientific research and practical value to explore the function of transcription factors and gene expression regulation.