37 results about "Response element" patented technology

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 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 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).

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.

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.

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.

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 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 invention belongs to the field of biological medicine, and relates to a lung poison cell screening model and construction and application thereof. The poison cell model provided by the invention contains a PPAR gamma promoter gene segment, a PPRE response element promoter gene segment and a reporter gene segment. The cell model can more sensitively and rapidly detect unknown toxicants with lung toxicity and also can perform early evaluation on whether drugs have side effects such as lung toxicity and the like, and has a good application prospect.

This invention discloses a rose salt tolerance gene RrSPL13, its miRNA resistance target rRrSPL13, and its applications. This invention belongs to the field of plant genetic engineering technology. The nucleotide sequence of the RrSPL13 gene is shown in SEQ ID NO.1, and its protein amino acid sequence is shown in SEQ ID NO.2. This invention uses large primer mutagenesis technology to synonymously mutate the miRNA response element of the RrSPL13 gene, obtaining the miRNA resistance target rRrSPL13, whose nucleotide sequence is shown in SEQ ID NO.3. Under salt stress, RrSPL13 is post-transcriptionally regulated by miRNA, while rRrSPL13 is not regulated. After isolating the RrSPL13 gene from wild rose, it was transformed into Arabidopsis thaliana and 'Purple Branch' rose. The salt tolerance of the transgenic Arabidopsis thaliana and 'Purple Branch' rose was significantly enhanced, indicating that the RrSPL13 gene is a positive regulator of salt tolerance and has important application value in the field of breeding cultivated roses with salt tolerance and stress resistance.

The invention discloses an intelligent cell strain as well as a construction method and application thereof, belongs to the technical field of biology, and aims to solve the limitation of single-point static monitoring, strong equipment dependence or insufficient biological safety in the existing detection technology. The method comprises the following steps: constructing a response element PCMV-GRLBD-NES-TetR by using a truncated receptor GRLBD, inserting the response element PCMV-GRLBD-NES-TetR and a report element SB-PTCE-SEAP into a genome of a host fine HEK-293T cell, and selecting a monoclonal cell responding to glucocorticoid; wherein the report protein in the report element can also use other target proteins besides alkaline phosphatase (SEAP), including but not limited to tyrosinase (Tyrosinase) and red fluorescent protein (tdTomato). The cell strain can be used for developing high-throughput screening of targeted mental stress drug molecules, and material reserve is provided for development of anti-depression and anti-anxiety drugs.

The invention provides a high-sensitivity in-vitro pyrogen detection method, which is characterized in that on the basis of an in-vitro pyrogen detection method (reporter gene method), a transgenic cell which expresses a Toll-like receptor and transfects an NF-kB reaction element and a luciferase gene is used for detecting a pyrogen, the pyrogen is combined with the Toll-like receptor on the surface of the cell to activate an NF-kB signal channel, and the NF-kB reaction element and the luciferase gene are converted into the NF-kB signal channel. Therefore, the expression of luciferase is activated.

Bivalent compounds composition comprises one or more of the bivalent compounds. The bivalent compound comprises a cyclic-AMP response element binding protein (CBP) and / or adenoviral EA binding protein of 300 kDa (P300) ligand (CBP / P3000 ligand) conjugated to a degradation tag. The method of using the bivalent compounds is treating certain disease in a subject in need thereof. The method of identifying such bivalent compounds is disclosed.

A nucleic acid for the controlled expression of a nucleic acid encoding a pro-apoptotic protein in an individual, including: a regulatory polynucleotide including a minimal promoter and at least one AARE (amino acid response element) nucleic acid, the regulatory polynucleotide being activated in an individual upon consumption of a diet deficient in at least one essential amino acid; and a nucleic acid encoding a pro-apoptotic protein, which is placed under the control of the regulatory polynucleotide.

The present invention belongs to the field of biotechnology, in particular the field of agricultural biotechnology. More specifically, the present invention relates to a method for precisely introducing environmental response elements by means of gene editing (e.g., prime editing) to regulate the distribution of carbon assimilates and thereby increase the yield of plants.

The present disclosure generally relates to a gene circuit comprising a response element engineered from NR4A1 that increases the activation of chimeric antigen receptors by the delivery of bioactive molecules. In particular, the present disclosure provides a nucleic acid construct comprising a response element operably linked to a nucleic acid sequence of interest that responds to the activation of CAR-T cells. The present disclosure also relates to vectors and cells comprising the gene circuit and the nucleic acid construct. Also provided are methods of inducing an immune response, methods of treating a subject, and methods for delivering a bioactive molecule by activated T cells having a chimeric antigen receptor and a response element construct of the present disclosure.

The present invention provides a system for drug-inducible expression of a polynucleotide comprising a) a first nucleic acid sequence comprising a first promoter inducible by said drug, wherein the first promoter is operably linked to said polynucleotide, wherein said first promotor comprises a binding site for a DNA binding domain, wherein said binding site comprises at least one responsive element that is recognized by said DNA binding domain (DBD), and b) a second nucleic acid sequence comprising a second promoter, wherein the second promoter is operably linked to a nucleic acid sequence encoding a synthetic transcription factor, wherein said synthetic transcription factor comprises i) an activation domain (AD), wherein said AD comprises the p65 activation domain of the human transcription factor NFκB or a functional variant thereof, ii) said DNA binding domain (DBD), wherein said DBD comprises or consists of 3 zinc finger domains, iii) a ligand-binding domain (LBD), wherein said LBD is a modified human estrogen receptor which is able to bind said drug, and wherein said ligand-binding domain (LBD) is positioned at the C-terminus of said synthetic transcription factor, and c) said drug, wherein said drug is tamoxifen or a metabolite of tamoxifen.

Oncolytic HSV vectors comprising NF-κB response elements or Oct-3 / 4-SOX2 response elements in the regulatory regions of viral genes that affect the efficiency of viral replication.

The present invention relates to small secreted reporter-peptides 15 to 150 amino-acids in length comprising a response element, activated by one or more transcription factors induced by the pharmacology active substance to be analyzed following its interaction with a specific intracellular or cell surface molecule, functionally linked to a response element, a TATA box, a signal peptide, anchor and detection sequences to which antibodies can be raised, and a poy-A tail. The anchor sequence may differ from one peptide to another or may be common to multiple secreted reporter-peptides such that all the peptides can be analyzed simultaneously by ELISA using a detection antibody labelled with for example HRP, or by the use of a commonly available immuno-detection platform such MSD, Gyros, AlphaLisa, or Biacore. The detection sequence is unique to each secreted reporter-peptide and may be labelled with a Sulfo-Tag that permits detection of the peptide on the MSD platform, or Alexa that permits detection on the Gyros platform, or digoxigenin that permits detection of on the PerkinElmer AlphaLISA platform, or left unlabeled for detection by SPR on a Biacore platform. The present invention provides i.a. a substantial improvement of cell-based assays for analysis using automated immune-detection platforms and allows simultaneous analysis of multiple analytes, multiple sampling from a single cell culture, and obviates the necessity to lyse cells and remove cell debris by centrifugation prior to analysis on an automated immuno-detection platform. The present invention also provides a means of increasing the dynamic range, sensitivity and reducing the cost of cell-based assays and can be applied to existing engineered cell lines, such as those containing a reporter-gene such as a luciferase reporter-gene obviating the necessity to extensively re-engineer cell lines containing multiple molecular constructs.

The invention belongs to the field of drug detection, and particularly discloses a method for detecting antibody-dependent cell-mediated cytotoxic biological activity of human immune globulin, which comprises the following steps: 1) adding a target cell suspension into a pore plate, respectively adding human immune globulin solutions with a series of concentrations for incubation, and finally adding an effector cell suspension for incubation; (2) detecting fluorescence of the incubated pore plate in the step (1), drawing a dose effect curve and calculating half effect concentration; the target cell is a CHO-K1 cell for expressing SARS-CoV-2 Spike protein, and the effector cell is a Jurkat cell which stably expresses CD16a on a cell membrane and is stably transfected with a firefly luciferase gene which is driven to express by an NFAT response element. The detection method disclosed by the invention can be used for accurately and reliably detecting the antibody-dependent cell-mediated cytotoxic biological activity of two different human immune globulin preparations, namely IVIG and SCIG.

The invention discloses a dehydration response element binding protein FtDREB02 as well as a coding gene and application thereof. The dehydration response element binding protein FtDREB02 and the coding gene thereof are separated from tartary buckwheat through molecular cloning, the amino acid sequence of the dehydration response element binding protein FtDREB02 is shown as SEQ ID No.2, and the nucleotide sequence of the coding gene of the dehydration response element binding protein FtDREB02 is shown as SEQ ID No.1. Genetic transformation experiments prove that the encoding gene of the dehydration response element binding protein FtDREB02 can remarkably improve the plant drought tolerance, has the dual functions of enhancing the promoter activity of the anthocyanin synthase gene, and has application prospects in the aspects of improving the plant drought stress resistance or regulating the plant anthocyanin biosynthesis and the like.

An adenovirus or adenoviral vector is described that includes a non-native nucleotide sequence capable of expressing a chimeric protein comprising an N-terminal nucleotide binding domain of transactivation response element DNA-binding protein (TDP-43), a C-terminal domain derived from a splicing repressor, and an autoregulatory element. Methods of using the adenovirus or adenoviral vector to treat degenerative diseases such as inclusion body myocytosis, amyotrophic lateral sclerosis, and frontotemporal dementia are also described.