201 results about "Transcriptional Activator" patented technology

Methods are described for detecting protein-protein interactions, among two populations of proteins, each having a complexity of at least 1,000. For example, proteins are fused either to the DNA-binding domain of a transcriptional activator or to the activation domain of a transcriptional activator. Two yeast strains, of the opposite mating type and carrying one type each of the fusion proteins are mated together. Productive interactions between the two halves due to protein-protein interactions lead to the reconstitution of the transcriptional activator, which in turn leads to the activation of a reporter gene containing a binding site for the DNA-binding domain. This analysis can be carried out for two or more populations of proteins. The differences in the genes encoding the proteins involved in the protein-protein interactions are characterized, thus leading to the identification of specific protein-protein interactions, and the genes encoding the interacting proteins, relevant to a particular tissue, stage or disease. Furthermore, inhibitors that interfere with these protein-protein interactions are identified by their ability to inactivate a reporter gene. The screening for such inhibitors can be in a multiplexed format where a set of inhibitors will be screened against a library of interactors. Further, information-processing methods and systems are described. These methods and systems provide for identification of the genes coding for detected interacting proteins, for assembling a unified database of protein-protein interaction data, and for processing this unified database to obtain protein interaction domain and protein pathway information.

Compositions, methods, strategies, kits, and systems for the supercharged protein-mediated delivery of functional effector proteins into cells in vivo, ex vivo, or in vitro are provided. Compositions, methods, strategies, kits, and systems for delivery of functional effector proteins using cationic lipids and cationic polymers are also provided. Functional effector proteins include, without limitation, transcriptional modulators (e.g., repressors or activators), recombinases, nucleases (e.g., RNA-programmable nucleases, such as Cas9 proteins; TALE nuclease, and zinc finger nucleases), deaminases, and other gene modifying / editing enzymes. Functional effector proteins include TALE effector proteins, e.g., TALE transcriptional activators and repressors, as well as TALE nucleases. Compositions, methods, strategies, and systems for the delivery of functional effector proteins into cells is useful for therapeutic and research purposes, including, but not limited to, the targeted manipulation of a gene associated with disease, the modulation of the expression level of a gene associated with disease, and the programming of cell fate.

Disclosed herein are in vitro assays for the identification of interactions between proteins or other molecules, the identification of transcriptional activator proteins, and the detection of compounds that inhibit protein / protein or protein / compound interactions. Also disclosed herein are in vitro assays for the selection of interacting proteins and transcriptional activator proteins out of libraries.

Engineered transcriptional activator-like effectors (TALEs) are versatile tools for genome manipulation with applications in research and clinical contexts. One current drawback of TALEs is their tendency to bind and cleave off-target sequence, which hampers their clinical application and renders applications requiring high-fidelity binding unfeasible. This disclosure provides engineered TALE domains and TALEs comprising such engineered domains, e.g., TALE nucleases (TALENs), TALE transcriptional activators, TALE transcriptional repressors, and TALE epigenetic modification enzymes, with improved specificity and methods for generating and using such TALEs.

Engineered transcriptional activator-like effectors (TALEs) are versatile tools for genome manipulation with applications in research and clinical contexts. One current drawback of TALEs is their tendency to bind and cleave off-target sequence, which hampers their clinical application and renders applications requiring high-fidelity binding unfeasible. This disclosure provides engineered TALE domains and TALEs comprising such engineered domains, e.g., TALE nucleases (TALENs), TALE transcriptional activators, TALE transcriptional repressors, and TALE epigenetic modification enzymes, with improved specificity and methods for generating and using such TALEs.

Increasing tolerance to butanol in yeast has been accomplished by increasing activity of the filamentous growth response. Yeast with increased expression of MSS11p, a transcriptional activator of the filamentous growth response pathway had increased tolerance to isobutanol. These yeast may be used for improved butanol production.

Engineered transcriptional activator-like effectors (TALEs) are versatile tools for genome manipulation with applications in research and clinical contexts. One current drawback of TALEs is that the 5′ nucleotide of the target is specific for thymine (T). TALE domains with alternative 5′ nucleotide specificities could expand the scope of DNA target sequences that can be bound by TALEs. Another drawback of TALEs is their tendency to bind and cleave off-target sequence, which hampers their clinical application and renders applications requiring high-fidelity binding unfeasible. This disclosure provides methods and strategies for the continuous evolution of proteins comprising DNA-binding domains, e.g., TALE domains. In some aspects, this disclosure provides methods and strategies for evolving such proteins under positive selection for a desired DNA-binding activity and / or under negative selection against one or more undesired (e.g., off-target) DNA-binding activities. Some aspects of this disclosure provide engineered TALE domains and TALEs comprising such engineered domains, e.g., TALE nucleases (TALENs), TALE transcriptional activators, TALE transcriptional repressors, and TALE epigenetic modification enzymes, with altered 5′ nucleotide specificities of target sequences. Engineered TALEs that target ATM with greater specificity are also provided.

Disclosed is a method using an alkane response element (ARE) from, e.g., Acinetobacter spp. to (i) identify and clone hydrocarbon biosynthesis genes, (ii) identify and clone hydrocarbon transporter genes (iii) identify and clone hydrocarbon response genes. Screening cells were developed that expressed a transcriptional activator, e.g., alkR, and included a reporter gene, e.g., GFP operatively linked to an ARE promoter, e.g., the alkM promoter. The cells were transformed with libraries from organisms capable of hydrocarbon biosynthesis. Transformed cells that expressed the reporter gene harbored library-derived genes involved in one or more of the above-mentioned processes; and these genes were isolated from the cells using standard molecular biology techniques. Additional systems were designed wherein screening cells also expressed a gene identified in the original screen, e.g., an additional hydrocarbon pathway gene, e.g., an enhancer.

The invention relates to novel mammalian perixosome proliferator-activated receptor (PPAR) poplypeptides and their use. In a particular embodiment, PPAR polypeptides with mutations in the P-box domain possess advantageous properties as transcriptional activators for PPRE-bearing expression vectors and expression systems. The invention includes PPAR polypeptides, nucleic acids encoding them, expression systems, vectors, and methods for inducibly expressing a gene of interest. The methods and expression systems of the invetion provide improved dose-response or inducibility characteristics, improved and / or altered effects on cellular PPAR function, and / or improved transcriptional control. The selection of a homologous PPAR polypeptide to prepare the novel PPAR polypeptide of the invetion for a cell or tissue also improves the immune reaction side effect potential for particular uses.

The present invention relates to methods for treating and / or preventing cancer. In particular the present invention relates to ex vivo immunotherapeutic methods. The methods comprise decreasing Stat3 (signal transducer and activator of transcription3) expression and / or function in tumor cells and the administration of such cells to a subject in need of treatment and / or prevention. Other methods of the invention comprise activating T-cells by co-culturing the T-cells with the tumor cells with decreased Stat3 expression or function. The invention further encompasses methods comprising decreasing Stat3 expression or function in antigen-presenting cells and co-administering tumor cells and the antigen-presenting cells with decreased Stat3 function to a patient. The invention further relates to methods for stimulating dendritic cell differentiation.

The present invention provides for a gene obtained during the induction of microspore embryogenisis. The protein encoded by this gene renders plant cells embryongenic, and increases the regenerative capacity of the plant cell. Also disclosed is the regulatory region of this gene and its use for directing the expression of a gene of interest within a suitable host cell.

Engineered transcriptional activator-like effectors (TALEs) are versatile tools for genome manipulation with applications in research and clinical contexts. One current drawback of TALEs is that the 5′ nucleotide of the target is specific for thymine (T). TALE domains with alternative 5′ nucleotide specificities could expand the scope of DNA target sequences that can be bound by TALEs. This disclosure provides methods and strategies for the continuous evolution of proteins comprising DNA-binding domains, e.g., TALE domains. In some aspects, this disclosure provides methods and strategies for evolving such proteins under positive selection for a desired DNA-binding activity and / or under negative selection against one or more undesired (e.g., off-target) DNA-binding activities. Some aspects of this disclosure provide engineered TALE domains and TALEs comprising such engineered domains, e.g., TALE nucleases (TALENs), TALE transcriptional activators, TALE transcriptional repressors, and TALE epigenetic modification enzymes, with altered 5′ nucleotide specificities of target sequences. Engineered TALEs that target ATM with greater specificity are also provided.

A method for regulating expression of a tet operator-linked gene in a cell of a subject is disclosed. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. Alternatively, in another embodiment, the method involves obtaining the cell from the subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding a tTA, to form a modified cell, administering the modified cell to the subject, and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecule can be within a single molecule (e.g., in the same vector) or on separate molecules.

The present invention provides compositions and methods for regulating expression of nucleotide sequences in a plant. Compositions comprise novel nucleic acid sequences encoding a transcription factor involved in modulating gene expression in response to abiotic stress such as cold or drought. Methods for expressing the nucleic acid sequence in a plant and improving cold and / or drought tolerance of plants are also provided.

The present invention provides a panel of transcriptional activator fusion proteins which comprises both tetracycline controlled transactivator proteins and reverse tetracycline transactivator proteins. These transactivators have novel phenotypes such as altered basal transcriptional activity in the absence of doxycycline, altered induced transcriptional activity in the presence of doxycycline, or differential induction by tetracycline and analogs of tetracycline.

The present invention provides a pair of short peptides, proteins and polynucleotides, a host cell and application thereof, and provides propose an artificial synthetic method for transcriptional activator-like effector nuclease (TALEN) gene of human cell endogenous IL2RG gene efficient targeting, and a method for orientation targeting by using a recombinant plasmid containing the gene. The TALEN comprises a pair of transcription activator-like effector (TALEs) proteins and DNA incision enzyme catalytic subunit respectively fused with the proteins, and can respectively recognize two adjacent sites on human IL2RG gene exon 2. On the basis of designing a TALEs amino acid sequence, a nucleotide sequence encoding the TALEN is synthesized and a vector containing the nucleotide sequence is constructed. By using the TALEN plasmid to transfect cells, the efficiency of cell targeting can be greatly improved.

Compositions and methods for modulating the production and characteristics of oil in a plant or plant part thereof are provided. Compositions of the invention are nucleotide constructs that provide for expression of transcriptional activators that increase lipid biosynthesis in a plant in combination with the inhibition or disruption of starch biosynthesis and / or starch storage, and optionally in combination with expression of another polynucleotide of interest. Compositions also include transformed plants, plant cells, plant tissues, and plant seeds comprising these nucleotide constructs. The methods of the invention comprise introducing a combination of polynucleotides into a plant, wherein the polynucleotides provide for increased lipid biosynthesis and disruption of starch biosynthesis and / or starch storage, and optionally alteration of fatty acid metabolism. The methods and constructs find use in alteration of oil phenotype in a plant or plant part thereof.

The invention relates to a high-efficiency artificial activating transcription factor dCas9-TV, and a coding gene and applications thereof. According to a construction method, the carboxyl terminal ofnuclease inactivated Cas9 protein (dCas9) is connected with a plurality of copies of VP64 and TAL transcription-activating domains so as to obtain a series of novel artificial activating transcription factors, and obtain dCas9-TV with the best transcriptional activation activity via screening. When only one guide RNA (gRNA) is adopted for targeting a specific gene promoter, dCas9-TV is capable ofrealizing high efficiency activating of transcription of endogenous genes of Arabidopis thaliana and paddy rice; when a plurality of gRNA are adopted for targeting a plurality of target genes, dCas0-TV is capable of realizing transcription activation of a plurality of genes. In addition, it is confirmed that dCas9-TV possesses the same high efficiency targeting transcription activation activity in human cells. An in vitro assembled dCas9-TV / gRNA ribonucleoprotein compound can be adopted for transcription activation of Arabidopis thaliana and paddy rice endogenous genes. The high-efficiency artificial activating transcription factor dCas9-TV can be adopted in the fields such as genome genetic screening, metabolite biosynthesis pathway reconstruction, and crop improvement.

The present invention provides compositions and methods for regulating expression of nucleotide sequences in a plant. Compositions comprise novel nucleic acid sequences encoding a transcription factor involved in modulating gene expression in response to abiotic stress such as cold or drought. Methods for expressing the nucleic acid sequence in a plant and improving cold and / or drought tolerance of plants are also provided.

In this invention, a method is described that allows for the efficient creation and identification of validated biological materials that greatly enhance the ability to perform polysome-mediated RNA profiling, such as constitutive, cell type-, tissue type-, or condition-enhanced RNA profiling. The method relies on the use of a tri-partite plant binary expression vector comprised of the following components: a) a DNA promoter element that drives expression of a sequence specific transcription activator protein such as a LexA:Gal4 fusion protein in a unique desired pattern, b) a DNA promoter element comprising a target site for the transcriptional activator protein, such as opLexA, fused to a nucleotide encoding an epitope tagged ribosomal component protein and c) a DNA promoter element comprising a target site for the transcriptional activator protein, such as opLexA, fused to a nucleotide encoding an in vivo reporter protein. By visualization of the co-regulated reporter, this method allows for in planta confirmation that the promoter element is driving expression, such as constitutive, cell type-, tissue type-, or condition-enhanced expression, of the tagged ribosomal protein in the desired cell or tissue types.

Increasing tolerance to butanol in yeast has been accomplished by increasing activity of the filamentous growth response. Yeast with increased expression of MSS11p, a transcriptional activator of the filamentous growth response pathway had increased tolerance to isobutanol. These yeast may be used for improved butanol production.

The present invention provides compositions and methods for regulating expression of nucleotide sequences in a plant. Compositions comprise novel nucleic acid sequences encoding a transcription factor involved in modulating gene expression in response to abiotic stress such as cold or drought. Methods for expressing the nucleic acid sequence in a plant and improving cold and / or drought tolerance of plants are also provided.

The invention aims to provide a bacillus licheniformis and application thereof. The preservation number of a bacterial strain of the bacillus (Bacillus lincheniformis) LV005 is CCTCC M 2013262. The bacillus licheniformis LV005 disclosed by the invention is screened from an alimentary canal of a prawn, and has the advantages of improving the composition and the number of dominant microbial populations in the alimentary canal of the prawn, forming and maintaining good microecological balance of the alimentary canal, and resisting the adhesion and the proliferation of viruses in the alimentary canal through nutrition competition, space competition, bacteriocin secretion and the like. The bacterial strain can activate the expression of prophenoloxidase, superoxide dismutase, lysozyme, heat shock protein, lipopolysaccharide-glucan conjugated protein, signal transduction, transcriptional activation factors and the like of the prawn with relevant pathogenic infection resistant molecules, and retard the proliferation rate of the viruses in the body of the prawn, so that the bacterial strain has various biological functions of improving the activity of the protease, the lipase and the amylase of the prawn, promoting the digestion capacity of the prawn and the digestion for feed nutrients, improving the stress response resistant capacity of the prawn and the like.

The present invention provides an expression system, the system comprising: a first nucleic acid molecule having a cell specific promoter; a second nucleic acid molecule encoding a transcriptional activator; a third nucleic acid molecule having a first recognition sequence of the transcriptional activator; a fourth nucleic acid molecule having a first promoter and a first regulatory element; a fifth nucleic acid molecule encoding a first regulatory protein; a sixth nucleic acid molecule having a second recognition sequence of the transcriptional activator; a seventh nucleic acid molecule having a second promoter and a second regulatory element; an eighth nucleic acid molecule encoding a second regulatory protein; and: a ninth nucleic acid molecule configured to conditionally inhibit expression of the first regulatory protein; and a tenth nucleic acid molecule configured to conditionally inhibit expression of the second regulatory protein, wherein the first regulatory element is adaptedto inhibit the function of the first promoter by binding to the second regulatory protein, and the second regulatory element is adapted to inhibit the function of the second promoter by binding to the first regulatory protein.