53 results about "Riboswitch" patented technology

The invention discloses a gene control device and method based on CRISPR-Cas9. The device comprises a nucleotide sequence, wherein the nucleotide sequence is an sgRNA itself or can be transcribed into an sgRNA, the sgRNA structurally comprises a first part bonded with inactivated Cas9 protein and a second part connected to the 3'end of the first part, the 5'end of the first part is provided with an antisense sequence, and the second part is an aptamer sequence part which comprises a ligand binding part and an aptamer stem part; when no ligand exists, the aptamer sequence is pair-bonded with the aptamer stem part; when a ligand exists, the aptamer sequence is dissociated from the aptamer stem part so as to be pair-bonded with a target DNA. By integrating an RNA riboswitch bonded with a specific biological signal into the sgRNA, a bridge of cell signal intensity and gene expression intensity is established, any target gene can be activated or silenced on the basis that biological signal identification is achieved, and artificial connection is built between signal ligands and cytogene.

The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and / or NK cells without prior ex vivo stimulation. The methods typically include engineered signaling polypeptides that can include a lymphoproliferative element, and / or a chimeric antigen receptor (CAR), for example a microenvironment restricted CAR. Additional elements of such engineered signaling polypeptides are provided herein, as well as vectors, such as retroviral vectors, packaging cell lines and methods of making the same. Furthermore, recombinant retroviruses and methods of making the same are provided. Numerous controls are provided, including riboswitches that are controlled, for example in vivo, by nucleoside analogues.

An intronic, self-splicing riboswitch is configured for enzyme-product specificity by introducing an appropriate aptamer. This then provides a sensing-expression construct, whereby the presence of an enzyme product in the cell triggers self-splicing of the intron sequence to restore the reading frame of the reporter gene and as such to drive expression of the gene product. The sensing construct expresses a protein which marks the cell or permits its growth or survival in or on an otherwise selective media. In this way, introduction or the presence of such product sensing-reporter constructs in cells can be harnessed to provide a multi-parallel rapid screening of cells or libraries for desirable enzyme variants.

The present invention relates to methods for constructing a recombinant fungal host cell comprising one or more copies of a polynucleotide construct integrated in its genome, said method comprising transforming a fungal host cell with an integrative polynucleotide construct comprising a first polynucleotide encoding a selectable marker, wherein the first polynucleotide, a 5′ untranslated region thereof and / or a riboswitch operably linked therewith comprises a spliceosomal intron which has 5 nucleotides or less between its branch site and its acceptor site; and a second polynucleotide encoding a polypeptide of interest; as well as suitable polynucleotide constructs, resulting fungal host cells and methods of manufacture.

The present invention relates novel flavin derivatives and other flavin derivatives, their use and compositions for use as riboswitch ligands and / or anti-infectives. The invention also provides method of making novel flavin derivatives.

The present invention relates to riboswitches that have been engineered to regulate pre-mRNA splicing. In particular, the insertion of a high affinity theophylline binding aptamer into the 3′ splice site region, 5′ splice site region, or branchpoint sequence (BPS) of a pre-mRNA modulates RNA splicing in the presence of theophylline. Accordingly, the aspects of the present invention include, but are not limited to, theophylline-dependent riboswitches which modulate RNA splicing, methods of modulating RNA splicing using theophylline and its corresponding riboswitches, methods of improving / identifying theophylline-dependent riboswitches, methods of treating diseases associated with or caused by abnormal RNA splicing.

The invention relates to a construction method and applications of bacillus subtilis with high yield of glucosamine. The construction method comprises the following steps: transforming a GFA1 gene coming from a brewer's yeast, a glms riboswitch gene and an EGFP enhanced fluorescent protein coding gene into bacillus subtilis, and carrying out construction to obtain bacillus subtilis with high yield of glucosamine. According to the invention, for the first time, the glms riboswitch gene and the EGFP gene are connected into PHT01, then a GLMS-EGFP sensor is constructed, the change of fluorescence intensity is shown by responding to the change of the amount of glucosamine 6-phosphate in bacillus subtilis cells, and through a flow cytometry, the synthesis condition of glucosamine can be known in real time.

The present invention relates to methods for constructing a recombinant fungal host cell comprising one or more copies of a polynucleotide construct integrated in its genome, said method comprising transforming a fungal host cell with an integrative polynucleotide construct comprising a first polynucleotide encoding a selectable marker, wherein the first polynucleotide, a 5′ untranslated region thereof and / or a riboswitch operably linked therewith comprises a spliceosomal intron which has 5 nucleotides or less between its branch site and its acceptor site; and a second polynucleotide encoding a polypeptide of interest; as well as suitable polynucleotide constructs, resulting fungal host cells and methods of manufacture.

The invention discloses a reporting system for detecting the cyclic diguanylic acid (c-di-GMP) content in living cells and an application of the reporting system. The reporting system comprises a promoter, a riboswitch, an SD (Shine-Dalgarnosequence) sequence and a reporter gene which are connected in sequence. The application is characterized in that the reporting system is transferred into cells to be detected, reconstitution cells are placed into a culture solution for culture until the OD600 (Optical Density) of the culture solution reaches 0.4 to 0.6, then the expression quantity of the reporter genes in the recombinant cells is detected, and the cyclic diguanylic acid content in the cells to be detected is calculated according to the following formulas: cyclic diguanylic acid content=-1072ln (miller unit-CK) +793.35, wherein the miller unit represents the expression quantity of the reporter genes in the recombinant cells; and the CK represents the background level expression quantity of the reporter genes in the cells to be detected. By adopting the reporting system disclosed by the invention, the c-di-GMP content in the living cells can be conveniently detected; the operation method is simple, convenient and easy to implement; and the detection result is real and reliable.

The invention discloses a method for detecting an affinity of a guanine riboswitch based on a graphene biosensor. The graphene biosensor is composed of a glass substrate and a graphene layer. Indium tin oxide (ITO) is arranged on two sides of the upper surface of the glass substrate. A part of the upper surface of the indium tin oxide on the two sides are covered by graphene layers, a part on oneside that is not covered by graphene is taken as a source electrode, and a part on the other side that is not covered by graphene is taken as a drain electrode. The method does not need to be marked,is simple to operate and convenient to use. High selectivity and specificity are achieved at a low use voltage, and the safety is good.

Provided herein are libraries of scaffolds derived from riboswitches and small ribozymes and their methods of use. The scaffolds of the invention yield aptamers that are easily identified and characterized by virtue of the structural scaffold. The nature of the scaffold predisposes these RNAs for coupling to readout domains to engineer biosensors that function in vitro and in vivo. Biosensors, synthetic RNA agents and synthetic DNA agents, and their methods of use, are also provided.

The invention uses Bacillus as a starting strain, uses a carbon metabolism response element cre in a maltose promoter to transform a target, a lysine ribose switch is fused at the same time, a mutantof the maltose promoter is constructed, the mutant that can reduce the leaky expression of the maltose promoter and improve the induced expression strength of the maltose promoter is obtained, and anexpression vector and host cell which contain the mutant of the promoter are constructed, so as to be conducive to the application of the maltose promoter in protein expression.

The present invention includes novel compounds and pharmaceutically acceptable formulations of said compounds which exhibit antibiotic activity against microorganisms bearing a guanine riboswitch that controls the expression of the guaA gene, including organisms which are resistant to certain antibiotic families, and which are useful as antibacterial agents for treatment or prophylaxis of bacterial infections in animals or in humans, in particular but not limited to infections of the mammary gland, or their use as antiseptics, agents for sterilization or disinfection.

The present invention is provides isolated riboswitches with FRET pairs for distinguishing changes in regulatory interactions controlled by the expression platform domain found in riboswitches. The invention further provides methods of using those riboswitches to detect structural changes in the expression platform domain and to identify potential antibiotics.

The present invention relates novel flavin derivatives and other flavin derivatives, their use and compositions for use as riboswitch ligands and / or anti-infectives. The invention also provides method of making novel flavin derivatives.

The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and / or NK cells without prior ex vivo stimulation. The methods typically include engineered signaling polypeptides that can include a lymphoproliferative element, and / or a chimeric antigen receptor (CAR), for example a microenvironment restricted CAR. Additional elements of such engineered signaling polypeptides are provided herein, as well as vectors, such as retroviral vectors, packaging cell lines and methods of making the same. Furthermore, recombinant retroviruses and methods of making the same are provided. Numerous controls are provided, including riboswitches that are controlled, for example in vivo, by nucleoside analogues.

The invention relates to a recombinant plasmid for promoting glutathione synthesis by dynamically regulating ATP, an engineering bacterium and an application thereof, and belongs to the technical field of fermentation. In the invention, a gene recombination technology is utilized; firstly, a riboswitch ydaO module, a ppk gene, a gshf gene segment and a plasmid vector are recombined to construct arecombinant plasmid; and then, the recombinant plasmid is transferred into an escherichia coli competent state, and an ATP regulation and control method based on the riboswitch ydaO module and polyphosphokinase (ppk) is constructed in escherichia coli so that an ATP concentration can be controlled at a certain level, and synthesis of intracellular ATP dependent products is facilitated.

The present invention relates novel flavin derivatives and other flavin derivatives, their use and compositions for use as riboswitch ligands and / or anti-infectives. The invention also provides method of making novel flavin derivatives.

The invention provides methods for treating or inhibiting infection by Clostridium difficile in a subject in need of such treatment, comprising administering an effective amount of a compound binding to a CD3299 riboswitch, as well as assays for identifying compounds useful in such treatment, and the use of particular compounds in such treatment.

The invention belongs to the field of biotechnology, relates to a novel antibiotic drug screening target spot, particularly to a sequence and a preparation method of aac riboswitches, and an application of the aac riboswitches as the novel target, antibiotic screening target. The invention provides a riboswitch, of which the DNA sequence is shown as SEQ IN NO 1, wherein N is any one of A, T, G or C; or the DNA sequence of the riboswitch is shown as SEQ IN NO 2. The characters of the riboswitch that the RNA sequence thereof can be combined with an antibiotic easily and further expression of a resistance gene is caused are utilized, so that the riboswitch can serve as a drug target for screening a novel antibiotic or reducing drug resistance of microorganisms. The riboswitch provides a new idea for an inductive drug resistance research.

The invention provides a recombinant adeno-associated virus vector capable of regulating and controlling genetic expression. Construction operation of the recombinant adeno-associated virus vector includes the following steps that riboswitch elements composed of theophylline aptamer and tabacco ringsport virus hammerhead ribozyme are recombined into a Kal gene sequence, a specific primer pair is designed to amplify the sequence according to the insertion position of the riboswitch elements in the Kal gene, and the rAAV vector is packaged; the mode of recombining the riboswitch element into the Kal gene sequence includes the steps that the single switch element is inserted into the 5'tail end and / or 3'tail end UTR of the Kal gene, or the multiple switch elements are inserted into the 5'tail end and / or 3'tail end UTR of the Kal gene in an end-to-end mode. The vector can regulate and control transgenosis expression in a mammal and has an obvious regulating and controlling function.

The invention discloses an aptamer screening method and application thereof. The method can be used for screening aptamers combined with small molecules. The screening method disclosed by the invention can be used for solving the problems of difficulty in experimental design, complex screening flows and the like in the existing SELEX technology, and more importantly, a RNA aptamer can be obtained by screening by using the method, is an aptamer enzyme on one hand, is an artificial riboswitch on the other hand, and can be applied to the 5'or 3 'end of a gene to directly regulate and control the expression of the gene. In a word, the method disclosed by the invention can be used for generating the aptamers combined with the small molecules, so that the aptamers can be applied to various types of detection, diagnosis assistance and the like, and can also be used for regulating and controlling gene expression as the artificial riboswitch.

The present disclosure identifies compounds that bind to a t-RNA-dependent riboswitch of aaRS gene expression unique to Gram-positive bacteria. The compounds have anti-bacterial activity.

The present invention relates to a regulatable gene expression construct comprising a nucleic acid molecule comprising two or more regulation sequences encoding respective RNA molecules comprising a riboswitch responsive to an effector compound, said riboswitch being operably linked to a respective coding region which encodes a respective modulator compound for modulating the action of a respective growth regulator compound and each said riboswitch in each regulation sequence being selected be responsive to the same effector compound to trigger expression of its respective modulator compound. The invention also relates to a method of using the regulatable gene expression construct for selecting from a metagenomic library a primary modulator compound which effects a chemical transformation of a substrate into said effector compound or transports said effector compound into a micro-organism comprising the regulatable gene expression construct.

Methods, compositions and assays that measure the effect of a test compound on induction of ligand-induced ribowitch-mediated transcription termination are disclosed. The methods and the assays are useful in identifying drug candidates that modulate transcription by binding to a riboswitch, for example.

The invention is a novel series of compounds represented by general formula, its analogs, tautomeric forms, stereoisomers, pharmaceutically acceptable salts and polymorphs wherein said compound described is a group of antisense molecules possessing immediate global cellular penetration due to their molecular structure and lipophilicity. The molecules are combinations of ethyl and methyl pyruvates. They induce riboswitch activity and reduce target RNA and nucleic acids such as HbA1c. They reduce inflammation and enhance energy production. These features improve therapeutic outcomes of diabetes, neuropathy and cellular aging.

The TPP riboswitch is a target for antibiotics, herbicides, algicides, fungicides and other utilities. The atomic structure of the binding pocket of the TPP riboswitch has been resolved. Compounds identified and optimized using this information can be used to stimulate, activate, inhibit and / or inactivate the TPP riboswitch.

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

The present invention provides bioengineered organisms producing elevated levels of thiamine and / or thiamine derivatives. Particularly, the present invention discloses that modifying TPP-responsive riboswitch results in accumulation of thiamine and / or its derivatives.

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are alsocontemplated.