65 results about "Response element" patented technology

A fusion protein comprising a DNA binding domain operably-linked to a dimerization domain, wherein the DNA binding domain specifically binds to a response element.

The invention relates to a biosensor capable of efficiently responding to vanillin, and belongs to the technical field of bioengineering. An aldehyde response transcription factor YqhC from Escherichia coli and a promoter element PyqhD regulated by the aldehyde response transcription factor YqhC are used for establishing a vanillin response RFP report system, a yqhC gene is subjected to semi-rational design, a mutation library is constructed through saturated mutation, vanillin is used as a substrate, and a non-background fluorescence response, a non-background fluorescence response function and a non-background fluorescence response function are obtained by screening response concentration and response threshold values. The YqhC mutant has a wider substrate detection range (0-10 mM) and higher response strength (167 times), and is named as YqhC-V18. The biosensor containing a vanillin response element (YqhC-V18) and a fluorescent protein report element (RFP) is further obtained, and can be applied to detection of a sample containing vanillin, or screening of a vanillin production strain, or regulation and control of the production process of vanillin.

The application relates to establishment and application of an isobutyraldehyde biosensor and belongs to the technical field of bioengineering. yqhD The RFP report system responding to isobutyraldehyde is established, the yqhC gene is rationally designed, a mutant library is constructed through random mutation by using error-prone PCR, isobutyraldehyde is used as a substrate, and through screening of response concentrations and response thresholds, a YqhC mutant without background fluorescence response, with a wider substrate detection range (0-5 g / L) and higher response intensity (24 times) is obtained, and the YqhC mutant is named YqhCm9. The isobutyraldehyde response element (YqhCm9) and the fluorescent protein report element (RFP) are used to construct a high-efficiency and specific isobutyraldehyde biosensor, real-time monitoring of an isobutyraldehyde production process and screening of an isobutyraldehyde production strain can be carried out.

The invention relates to a method for determining the biological activity of FGF21 (Fibroblast Growth Factor Receptor 21), which comprises the following steps: infecting and transfecting HEK293T cells by using an overexpression lentivirus containing FGFR1 and beta-Klotho genes and a reporter gene plasmid containing an SER reaction element so as to obtain a reporter gene-containing cell strain capable of stably expressing FGFR1, beta-Klotho and SRE; after FGF21 stimulation is added, the expression of a luciferase reporter gene at the downstream of the SER-containing promoter is activated; after co-incubation culture, adding a luciferase substrate, and fitting a four-parameter curve according to a measured reporter gene signal value, so as to determine the biological activity of the FGF21. According to the method provided by the invention, the test period is shortened to 24 hours, the detection sensitivity for FGF21 can reach a nanogram level and is improved by 1000 times compared with that of a traditional glucose oxidation method, and the result is consistent. The method provided by the invention can be used for evaluating the biological activity of FGF21 and analogues thereof as candidate drugs for treating metabolic diseases such as diabetes mellitus and NASH, and plays an important guiding role in quality control in research, development and production of FGF21 drugs.

The present disclosure generally relates to genetic circuits including response elements engineered from the NR4A1 that augment chimeric antigen receptor activation with the delivery of a bioactive molecule. Particularly, the disclosure provides nucleic acid constructs including response elements operably linked to a nucleic acid sequence of interest such that are responsive to activation of CAR-T cells. The disclosure also relates to vectors and cells including the genetic circuits and the nucleic acid constructs. Also provided are methods of inducing an immune response, methods of treatment of subjects, and methods for delivering a bioactive molecule by an activated T cell having a chimeric antigen receptor and the response element constructs of the disclosure.

The present invention relates to the field of plant genetic engineering technology, and in particular to a wheat cold-resistant gene TaERF7-like and its application. The present invention provides an application of a wheat ethylene response element binding protein family AP2 / EREBP transcription factor gene TaERF7-like in plant cold resistance and advancing the flowering time of plants. Plants transformed with this gene can tolerate low temperature stress and advance the flowering time. The nucleotide sequence of the gene and the amino acid sequence of the encoded protein are shown in SEQ ID NO.1 and SEQ ID NO.2, respectively. By overexpressing the gene in Arabidopsis thaliana, the cold resistance of the transgenic plants can be significantly improved, and the flowering time of the plants can be advanced. The TaERF7-like gene of the present invention can be used as a genetic resource for genetic improvement of crops and has broad application prospects.

The invention relates to the technical field of gene engineering detection, and particularly discloses a gene detection system for promoting andrographolide synthesis based on a biosensor, a key gene ApDof29 for regulating and controlling andrographolide synthesis is determined through whole genome analysis, gene expression profile research and co-expression network analysis, the core regulation and control effect of ApDof29 is verified, and the gene detection system for promoting andrographolide synthesis based on the biosensor is used for promoting andrographolide synthesis. The invention also develops a real-time dynamic monitoring system, and realizes non-invasive monitoring of andrographolide content accumulation, biosynthesis rate and metabolic intermediate concentration change by utilizing the fusion of a specific response element and a reporter gene. Through comprehensive calculation processing of an accumulation abnormal coefficient, a synthesis rate abnormal coefficient and a concentration change abnormal coefficient, the regulation and control effect of ApDof29 is dynamically evaluated, and a scientific basis is provided for subsequent gene modification.

The present disclosure provides DNA molecules, such as transfer plasmids, useful for manufacturing packaged retroviral vectors encoding a therapeutic protein such as chimeric antigen receptors (CARs) and T cell receptors (TCRs). The encoded retroviral transcript in the DNA molecule includes a transgene, an elongation factor 1-alpha 1 (EFla) promoter, without the EFla intron, operably linked to the transgene and a Rev response element (RRE).

The invention relates to a hypoxia response type synNotch-ODD CAR with dual-target recognition and application thereof, and belongs to the technical field of cell therapy. The invention provides an anti-tumor expression vector. The anti-tumor expression vector comprises nucleic acid sequences of a CD133-synNotchCAR (chimeric antigen receptor) and an EGFR (epidermal growth factor receptor)-hypoxia response element CAR (chimeric antigen receptor). The invention further provides a synNotch-ODD CAR immune cell, and the synNotch-ODD CAR immune cell comprises the expression vector. The polypeptide can be activated and plays a role in killing tumor cells only under the conditions of hypoxia and simultaneous expression of EGFR and CD133 antigens by target cells, and the on-target / off-tube risk caused by activation of metastatic cancer cells or circulating tumor cells of traditional synNotch CAR can be effectively solved.

To provide contiguous DNA sequences encoding highly compact multi-input genetic logic gates for precise in vivo cell targeting, and methods of treating disease using a combination of in vivo delivery and such contiguous DNA sequences.SOLUTION: A contiguous polynucleic acid molecule encoding at least two cassettes is provided, where each cassette comprises a regulatory component and a response component.SELECTED DRAWING: None

The invention belongs to the technical field of biological engineering, and discloses a preparation method and application of Escherichia coli regulated by blue light, and the preparation method comprises the steps of blue light response element construction, strain engineering transformation, fermentation system design, culture condition optimization and product purification work.The blue light response element construction comprises the steps that a, a core light sensing assembly is selected; b, designing and synthesizing an EL222 coding sequence, c, constructing and synthesizing a promoter, and d, optimizing an EL222 binding site; the engineering modification of the strain comprises the following steps: a, integrating a photosensitive promoter and a target gene, and b, optimizing the expression level of EL222 protein. According to the invention, directional regulation and control of metabolic pathways can be realized at a subcellular level through the blue light response element, and through accurate control of illumination intensity and time, metabolic flux can be dynamically adjusted according to requirements, accumulation of metabolic intermediates is reduced, cytotoxicity is reduced, purity of a target product is improved, and the method is suitable for industrial production. The yield and the quality of a target product are improved through precise space-time regulation and control on escherichia coli metabolism.

In one aspect, the disclosure relates to inhibition of ring AMP-response element binding protein (CREB). The present disclosure relates to solid and salt forms of inhibitors of CBP / p300 family bromodomains consisting of salts and crystalline forms of formula (I). Compounds may be used to treat diseases or conditions associated with inhibition of the CBP / p300 family bromodomains. For example, the disclosure relates to compounds and compositions for inhibiting the CBP / p300 family bromodomains, methods of treating diseases or conditions associated with inhibition of the CBP / p300 family bromodomains (e.g., certain forms of cancer), and methods of synthesizing these compounds.

Described herein are switchable core promoters that may selectively promote transcription initiation in the presence of an activated response element, such as an activated enhancer. These switchable core promoters may be paired with cell type- or cell state-specific response element to produce engineered promoters that selectively promote transcription of a payload sequence in a target cell type or target cell state. Also described herein are methods of using switchable core promoters and polynucleotides containing switchable core promoters to selectively express a payload in a target cell type or target cell state.

The invention relates to the technical field of gene engineering detection, and particularly discloses a gene detection system for promoting andrographolide synthesis based on a biosensor, a key gene ApDof29 for regulating and controlling andrographolide synthesis is determined through whole genome analysis, gene expression profile research and co-expression network analysis, the core regulation and control effect of ApDof29 is verified, and the gene detection system for promoting andrographolide synthesis based on the biosensor is used for promoting andrographolide synthesis. The invention also develops a real-time dynamic monitoring system, and realizes non-invasive monitoring of andrographolide content accumulation, biosynthesis rate and metabolic intermediate concentration change by utilizing the fusion of a specific response element and a reporter gene. Through comprehensive calculation processing of an accumulation abnormal coefficient, a synthesis rate abnormal coefficient and a concentration change abnormal coefficient, the regulation and control effect of ApDof29 is dynamically evaluated, and a scientific basis is provided for subsequent gene modification.

The invention discloses a gene editing method for regulating and controlling the nitrogen utilization rate of rice and a regenerated rice plant, and relates to the technical field of agricultural biology. The gene editing method for regulating and controlling the nitrogen utilization rate of the rice comprises the following steps: selecting a key gene of nitrogen metabolism of the rice as a target gene; determining a target spot in a promoter region of the target gene; constructing a repair donor template containing a nitrate response element based on the sequences on the two sides of the target spot; wherein the nitrate response element is derived from a promoter region of rice nitrite reductase, and can respond to a nitrate signal to enhance the expression of a downstream gene; jointly delivering the repaired donor template and a CRISPR / Cas9 carrier to the rice callus through gene editing, and culturing; after the culture is completed, screening to obtain a regenerated rice plant with the nitrate response element accurately inserted in the genome. According to the application, the nitrate response element is inserted into the promoter region of the nitrogen metabolism key gene to activate gene expression induced by nitrate, so that nitrogen utilization of rice is enhanced, and regenerated rice plants with high nitrogen utilization efficiency are obtained.

The invention belongs to the field of gene engineering, and particularly relates to a large yellow croaker SOCS1 gene promoter and application thereof. The nucleotide sequence of the large yellow croaker SOCS1 gene promoter is as shown in SEQ ID NO. 1. The large yellow croaker SOCS1 gene promoter is constructed to a pGL3.0 vector and transfected to a carp epithelioma cell (EPC), so that the large yellow croaker SOCS1 gene promoter has relatively high basic activity, and the activity of the large yellow croaker SOCS1 gene promoter is remarkably reduced by a mutated interferon stimulus response element (ISRE) motif; under the stimulation of a virus analogue poly (I: C), the activity of the large yellow croaker SOCS1 gene promoter is further improved, and the activity of a mutated promoter is reduced. The large yellow croaker SOCS1 gene promoter provided by the invention can be used for promoting target gene expression, researching a fish antiviral immune mechanism and breeding fish transgenosis.

Provided herein are compositions and methods to treat a disease or condition associated with haplo-insufficient genes comprising administering to a subject in need thereof a dCas protein and one or more gRNA, wherein the one or more gRNA is complementary to a miRNA response element (MRE) on target mRNA expressed from the functioning allele of the haplo-insufficient gene so as to cause dCas / gRNA binding to the MRE thereby blocking miRNA repression of the functioning allele.

A method for detecting intercellular interaction, the method comprising: (1) preparing a first cell expressing a TCR activation response element and a reporter molecule, the first cell expressing TCR; (2) incubating the antigen peptide and a second cell expressing MHC to obtain a second cell loaded with the antigen peptide; (3) marking the first cell by using a first tag to obtain a first cell carrying the tag, and marking the second cell loaded with the antigen peptide by using a second tag to obtain a second cell carrying the tag; (4) wrapping the first cells carrying the labels, the second cells carrying the labels and the magnetic beads with an oil phase to prepare a plurality of liquid drops; (5) injecting a cell lysis solution into the single liquid drop, and sequencing the nucleic acid after lysis; and (6) determining whether the first cell and the second cell in the single liquid drop interact or not according to a sequencing result.

The invention belongs to the technical field of medical chemistry, and discloses an aza aromatic amine protein degradation targeting chimera (PROTAC) compound with the activity of degrading nuclear receptor co-inhibitor 1 (NCoR1) in a targeting manner, and a preparation method and application of the aza aromatic amine protein degradation targeting chimera (PROTAC) compound. The aza aromatic amine compound is prepared through acylation and N-alkylation reaction, and the structure of the compound is shown as a formula I to a formula IV. A Liver X Receptor Response Element-Luciferase reporter gene system (LXRE-Luc luciferase reporter gene system) determination experiment shows that when the compound is 10 [mu] M, the compound has an obvious liver receptor (LXR) regulation effect, and the compound has an obvious liver receptor regulation effect when the compound is 10 [mu] M; western blot experiment research shows that the compound has a remarkable degradation effect on NCoR1, and the compound XH-10 with the highest activity can generate a remarkable NCoR1 degradation effect when the concentration is 5 mu M. The compound is PROTAC with NCoR1 targeted degradation activity, and has the advantages of high activity and simple preparation process. The compound has an important value in the aspect of glycometabolism and ester metabolism regulation medicine development.

Described herein are switchable core promoters that may selectively promote transcription initiation in the presence of an activated response element, such as an activated enhancer. These switchable core promoters may be paired with cell type- or cell state-specific response element to produce engineered promoters that selectively promote transcription of a payload sequence in a target cell type or target cell state. Also described herein are methods of using switchable core promoters and polynucleotides containing switchable core promoters to selectively express a payload in a target cell type or target cell state.

Provided herein are nucleic acid constructs comprising one or more Nuclear Factor of Activated T-cells (NFAT) response elements and an expressible nucleic acid sequence encoding a first engineered polypeptide monomer, wherein the first engineered polypeptide monomer comprises a regulatable polypeptide payload and at least one drug response domain (DRD). The abundance, availability, and / or biological activity of the payload is regulated by interaction of an effective amount of an immune cell stimulator with the one or more NFAT response elements and by interaction of an effective amount of a ligand with the one or more DRDs.

The invention discloses a mitochondrial transcriptional regulatory factor Mt-CREB and application thereof in screening and preparing a drug for resisting brain aging. According to the present invention, based on the characteristic that the mitochondrial circular DNA has the conservative cAMP reaction element sequence CRE, the CREBWT capable of selectively identifying and combining the CRE or the amino acid RRPSYR mutant CREBDIEDML of the CREBWT at the 130-135th site is adopted as the key element of the gene enhancement regulation, and the CREBWT or the CREBDIEDML is selectively positioned to the mitochondria by using the mitochondrial targeting sequence so as to achieve the detection and the regulation of the mitochondrial gene expression enhancement; on one hand, the invention discloses an amino acid sequence of the Mt-CREBDIEDML, a nucleotide sequence for coding the Mt-CREBDIEDML, an expression vector containing the nucleotide sequence, and an application of the Mt-CREBDIEDML in preparation of a drug for resisting brain aging; on the other hand, the invention discloses a fluorescence and / or chemiluminiscence detection method for screening the anti-brain aging drugs by using the Mt-CREBWT. Through research on biochemical indexes, immunohistochemistry, gene expression level and the like, a new way is provided for preparing and screening anti-brain aging drugs and treating memory decline caused by aging.

The present disclosure provides high-performance immune-activation reporter cell line set. A set of nucleic acid constructs comprising one or more response elements operatively linked to a TATA box motif promoter or a minimal IL-2 promoter are provided. A set of reporter cell lines comprising a polynucleotide sequence encoding a reporter protein under the control of one or more response elements operatively linked to a TATA box motif promoter or a minimal IL-2 promoter is also provided. The provided set of reporter cells enable sensitive and signal-enhanced bioassay performance to reflect and evaluate adaptive immune cell activation / inhibition regulated by the immune receptors CD3, co-stimulatory CD28 and related signaling pathways, and avoid responding bias triggered by particular transcription factor. A controllable engineer method for a functional cell line is further provided.

The present disclosure provides a nucleic acid comprising a response element operably linked to a reporter nucleotide sequence. In various embodiments, the response element comprises 2 to 10 copies of a Z1 transcription factor binding site. Also provided is a cell comprising the nucleic acid. In various aspects, the cells stably overexpress adeno-associated virus receptors. The present disclosure further provides a method of determining the potency of a sample comprising an AAV vector.

Nucleic acid constructs comprising one or more activated T-cell nuclear factor (ATNF) response elements and an expressible nucleic acid sequence encoding a first manipulated polypeptide monomer are provided herein, wherein the first manipulated polypeptide monomer comprises a tunable polypeptide payload and at least one drug-sensitive domain (DSD). The abundance, availability, and / or biological activity of the payload are regulated by the interaction of an effective amount of immune cell-stimulating agent with the one or more ATNF response elements and by the interaction of an effective amount of ligand with the one or more DSDs.

The invention relates to a cat omega interferon biological activity detection method and a kit. According to the method, a stable reporter cell line is established by constructing a recombinant plasmid containing a cat omega-interferon specific interferon stimulus response element (ISRE) and a reporter gene (such as luciferase) and transfecting CRFK cells. During detection, report cells and a sample to be detected are co-incubated, cat omega-interferon induces report gene expression by activating a JAK-STAT-ISRE signal channel, the expression level is in direct proportion to interferon activity, and the activity can be quantified by detecting a luminescence value (RLU). The method overcomes the defects of long period (3-5 days), complicated operation and subjective result judgment of the traditional cytopathic effect inhibition method, has the advantages of rapidness (about 24 hours), safety (no live virus), objectiveness, accuracy, good repeatability and easiness in high throughput, and is suitable for quality control analysis of cat omega-interferon products.

This invention relates to the field of gene editing technology, specifically to a gene circuit-based specific gene expression system and module, a pharmaceutical composition, and its applications. The system includes a first vector and a second vector. The first vector includes a first expression cassette containing a cell-specific promoter, a coding sequence encoding a transcriptionally activated fusion protein, and a regulatory region sequence downstream of the coding sequence. The regulatory region sequence is configured to form a response element in the 3' untranslated region of the fusion protein's mRNA after transcription. This response element binds to a specific long non-coding RNA within the silenced cell, leading to the degradation of the fusion protein's mRNA. The second vector includes a second expression cassette containing an associated promoter that can be activated by the transcriptionally activated fusion protein, and a target gene downstream of the associated promoter. Advantages: This ensures that the target protein is expressed only in target cells and not in cancer cells, avoiding adverse effects on non-target cells or tissues, and reducing treatment risks and side effects.

Provided herein are nucleic acid constructs comprising one or more nuclear factor of activated T cells (NFAT) response elements and an expressible nucleic acid sequence encoding a first engineered polypeptide monomer, wherein the first engineered polypeptide monomer comprises a regulatable polypeptide payload and at least one drug responsive domain (DRD). The abundance, availability, and / or biological activity of the payload is regulated by interaction of an effective amount of immune cell stimulating agent with the one or more NFAT response element and by the interaction of an effective amount of ligand and the one or more DRDs.

The present invention provides a genetically modified non-human animal or the like that functionally lacks an endogenous ABCB1 gene and that functionally expresses the human ABCB1 gene, the genetically modified non-human animal or the like retaining a DNA construct for functionally expressing the human ABCB1 gene, wherein the DNA construct includes a coding sequence for the human ABCB1 gene, a CAR / PXR response element present in the 5' promoter region of the human ABCB1 gene, at least one intron of the human ABCB1 gene, and a sequence of the enhancer region present at the 3' end side of the human ABCB1 gene, and the DNA construct does not include at least a portion of the coding sequence for the human ABCB4 gene or includes only a portion of the human RUNDC3B gene.

The glucose-responsive transcription factor ChREBP (Carbohydrate Response Element Binding Protein) is a key mediator in the response to glucose in pancreatic β-cells, strongly regulating glycolytic and lipogenic pathways. Several protein regulators interact with ChREBP that are involved in its activation mechanism, including 14-3-3 protein. Small-molecules are provided, which are useful as stabilizers for ChREBP 14-3-3 complexes, which were found highly useful for medical treatments, i.a. in type 1 and type 2 diabetes.