107 results about "Repressor" patented technology

Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling the activity of proteins comprising a Cas9 variant fused to a functional effector domain, such as a nuclease, nickase, recombinase, deaminase, transcriptional activator, transcriptional repressor, or epigenetic modifying domain. For example, the inventive proteins provided comprise a ligand-dependent intein, the presence of which inhibits one or more activities of the protein (e.g., gRNA binding, enzymatic activity, target DNA binding). The binding of a ligand to the intein results in self-excision of the intein, restoring the activity of the protein.

The present disclosure relates to compositions and methods for reducing expression of MYC gene in a cell. In some embodiments, an expression repressor comprises a targeting moiety that binds a MYC promoter, anchor sequence, or super-enhancer. In some embodiments, the expression repressor comprises an effector moiety that represses transcription or methylates DNA. Systems comprising two expression repressors are also disclosed. The compositions can be used, for example, to treat cancers such as HCC.

The present disclosure provides Crispr / cas9-based repressors for silencing gene targets in vivo and methods of use.

A genetically modified yeast cell, wherein the yeast cell comprises a genetic modification comprising overexpression of yeast gene encoding porphobilinogen deaminase (HEM3), the HEM3 gene having at least 80% identity with SEQ ID No. 7. The genome of the modified yeast cell further comprises one or more genetic modifications in one or more genes selected from: genes coding for heme-dependent repressor of hypoxic genes (ROX1), genes coding for heme oxygenase (HMX1), genes coding for a receptor for vacuolar proteases (VPS10), and genes coding for vacuolar proteinase (PEP4), the one or more genetic modifications being such that expression of a polypeptide from such a gene is reduced or disrupted or the polypeptide expressed is non-functional.

The invention belongs to the technical field of biological medicines, and discloses an engineered bacterium for in-situ expression of a microtubule inhibition protein in a tumor and application of the engineered bacterium. The engineering bacteria are formed by constructing microtubule inhibition protein genes to an expression vector and finally transferring the expression vector into bacteria. The engineering bacterium can be colonized in a tumor microenvironment in a targeted manner, expresses microtubule inhibition protein, effectively inhibits tumor cells and remarkably inhibits solid tumor growth, and is excellent in safety.

The inventions provide mammalian cell lines integrated with recombinant adeno-associated virus (rAAV) plasmids, cell genomes and methods of making the cell genomes for production of rAAV. The inventions also provide regulatory fusion proteins (RFPs) (which can act as activators or repressors) and repressor proteins (which act as repressors), such as antibiotic repressors, and arranged operators to control transcription of at least one polynucleotide of interest, such as AAV Rep genes and / or non-AAV helper genes.

The present disclosure is directed to novel therapeutic approaches for the prevention, treatment and / or delaying progression of chronic injury, progressive loss of functional parenchymal cells, or fibrosis of an organ. Specifically disclosed are agents for use in increasing homodimer-formation of ARNT in an organ in the prevention, treatment and / or delaying progression of chronic injury, progressive loss of functional parenchymal cells, or fibrosis of said organ, as further defined in the claims. In embodiments, said agent may be (i) an inhibitor of protein phosphatase 2A (PP2A) activity, (ii) an inhibitor of the transcriptional repressor complex FKBP12 / YY1, or (iii) an expression construct, which is capable of over-expressing ARNT in said organ, as well as combinations of (i), (ii), and / or (iii).

A complex, comprising a first fusion and a second fusion, wherein one of the first fusion and the second fusion comprises a DNA methylation domain and at least one recruitment domain A, and the other fusion comprises a transcriptional repressor domain and at least one recruitment domain A′; and the recruitment domain A and the recruitment domain A′ interact with each other, so that one of the first fusion and the second fusion or a part thereof is recruited to the vicinity of the other fusion. Also provided are use of the complex in the preparation of a product for inhibiting target gene expression and the preparation of a drug.

This article provides gene repressor systems that can be used to repress genes, the gene repressor systems comprising repressor fusion proteins, such as repressor fusion proteins containing DNA-binding proteins, in some cases catalytically dead CRISPR proteins, and guide nucleic acids (gRNA). Methods for using such systems to repress gene transcription are also provided.

This invention discloses a genetically engineered bacterium for synthesizing carotenoids, its construction method, and its applications, belonging to the field of genetic engineering technology. The genetically engineered bacterium uses a carotenoid-producing yeast strain as the starting material. The gene ROX1, encoding a heme-dependent hypoxia gene repressor, is knocked out, while the expression of genes STB5 (encoding a transcription factor involved in NADPH regeneration), DID2 (encoding a class E protein in the vacuole protein sorting pathway), and VOA1 (encoding an endoplasmic reticulum protein playing a role in the assembly of the V0 region of V-ATPase) is upregulated. This invention promotes carotenoid synthesis and significantly increases carotenoid yield through combined regulation of gene expression outside the carotenoid synthesis pathway, demonstrating promising application prospects.

Provided herein are compositions and methods for treating an autoimmune disease or disorder in a subject. In some cases, the compositions and methods use chemical inducers of proximity (CIPs). In some cases, the CIPs include a first moiety having specific binding for a BTB domain-containing protein or a transcriptional repressor expressed in B cell subpopulations and a second moiety having specific binding for a modulator of gene regulation. In some cases, the BTB domain-containing protein comprises BCL6. In some cases, the transcriptional repressor expressed in B cell subpopulations is PRDM1 or IRF4. In some cases the modulator of gene regulation comprises BRD4, CDK9, and / or p300.

The disclosure features compositions or systems and uses thereof, comprising a first polynucleotide encoding a target molecule, optionally a second polynucleotide encoding an effector, repressor, or endonuclease molecule; optionally a recognition or cleavage site in the first or second polynucleotide, and optionally a repressor / effector binding site in the first polynucleotide. The disclosure also features compositions or systems and uses thereof, comprising a messenger RNA (mRNA) comprising (i) an open reading frame encoding a polypeptide, and (ii) at least six miR142 target sites.

The application relates to methods for identifying putative regulatory elements that regulates a gene, comprising: obtaining a measure of intrinsic activity of a plurality of genomic elements; obtaining a measure of proximity between each of the genomic elements and the gene; scoring a predicted impact of each of the genomic elements on the gene as a function of the measure of intrinsic activity and the measure of proximity, wherein a plurality of predicted impacts scored are ranked to identify at least one genomic element as a putative regulatory element that regulates the gene; and optionally, training, optimizing, and / or validating the scoring of predicted impact using experimental or computational data describing functional interactions between the genomic elements and the gene. The application also relates to methods for identification of transcriptional enhancers and repressors regulating a gene associated with an agricultural trait of interest in plants or a disease phenotype in mammalians.

The present disclosure relates to compositions and methods for reducing the expression of the MYC gene in cells. In some embodiments, the expression repressor comprises a targeting moiety that binds to the MYC promoter, an anchor sequence, or a super enhancer. In some embodiments, the expression repressor comprises an effector moiety that suppresses transcription or methylates DNA. A system comprising two expression repressors is also disclosed. The compositions can be used, for example, for the treatment of cancers such as HCC.

The invention belongs to the technical field of gene engineering, and particularly relates to application of CmIAA27 in regulation and control of lateral branch development of chrysanthemum. According to the invention, phenotypic analysis is carried out on a CmCMT2-RNAi transgenic 'Shenma' plant obtained at an earlier stage, and lateral branches of the CmCMT2-RNAi transgenic 'Shenma' are increased in a vegetative growth period. RNA-seq is carried out on the gene CmIAA27, it is found that the expression quantity of the auxin inhibitory signal key gene CmIAA27 is remarkably increased, McrBC and BSP experiments show that the CHH type methylation level on a CmIAA27 promoter in a transgenic line is remarkably lower than that of WT, and the expression of the plant lateral branch development key inhibitory gene CmBRC1 in the transgenic line is remarkably reduced. Y1H, LUC and EMSA experiments show that the CmCMT2 can be directly combined with a CmIAA27 promoter and regulate and control the CHH type methylation level in the region so as to influence the expression of the CmIAA27. Through transcription analysis of CmCMT2-RNAi transgenic Shenma and WT, a key factor CmIAA27 for downstream regulation and control of chrysanthemum lateral branch development is obtained, functional verification is carried out, and excellent gene reserve and a new strategy are provided for regulation and control of chrysanthemum lateral branches.

Provided are plasmids comprising transcriptional control elements, including transcription factor binding motifs, telomeric repeat motifs, transcription factor repressor binding motifs and promoters for tunable protein expression in target cells; wherein the plasmid for controlled transcription in a target cell type comprises a transcription factor binding motif for NFIA and the cell expresses NFIA. In one embodiment, the cell type is lung cells and the transcription factor is Nuclear Factor IA (NFIA).

The present invention discloses the construction and application of a screening system for lactose or its analog biosensors in Bacillus subtilis. The present invention designs a "dual-screening" biosensor repressor screening gene circuit based on the lactose operon and applicable to Bacillus subtilis: the DNA binding activity of the repressor is screened through a negative screening lethal circuit; the allosteric activation activity is screened through a positive screening fluorescence signal; and finally, a biosensor with the recognition activity of the target lactose analog is obtained. Using this screening platform, novel biosensors responsive to substances such as lactose, 2'-fucosyllactose, and 3-fucosyllactose can be obtained. Moreover, it has been verified that the biosensors constructed according to the screening results of the present invention have high detection sensitivity and specificity.

Novel gene repressor systems are provided that are useful in animal cells that include engineered repressors with altered ligand-binding domains (LBDs) and DNA-binding domains (DBDs), as well as mutated operator sequences. In one aspect, the systems enable independent, drug-inducible control of multiple genes within a single animal or animal cell line. In one aspect, the disclosure provides both tetracycline-responsive repressors and sulfonylurea (SU)-responsive repressors — e.g., variants that respond specifically to ethametsulfuron-methyl (Es) or chlorsulfuron (Cs). Mutations are introduced to either the LBD (to alter ligand specificity), DBD (to alter operator binding), or the operator sequence itself, resulting in orthogonal systems.

The invention relates to the technical field of gene engineering, in particular to a bidirectional promoter Para *, an araa * expression system, application and an engineering strain. The araa * expression system comprises the bidirectional promoter Para *, a repressor protein araR gene module is arranged on one side of the bidirectional promoter Para *, and a fluorescent protein gene module is arranged on the other side of the bidirectional promoter Para *. Wherein the repressor protein araR gene module comprises a repressor protein araR gene and a first RBS sequence corresponding to the repressor protein araR gene; and the fluorescent protein gene module comprises a fluorescent protein gene and a second RBS sequence corresponding to the fluorescent protein gene. By characterizing and designing an exogenous inducible promoter, an ara * expression system capable of realizing inducible expression in xanthomonas campestris is obtained, and a basic element is provided for construction of a xanthomonas campestris genetic engineering strain.

Provided is a precursor plasmid used for preparing a screening-tag-free plasmid, the precursor plasmid comprising: (1) a conditioned replication initiation site having a plasmid replication initiation capacity that is dependent on regulatory proteins, wherein the conditioned replication initiation site has a first replication initiation state in the presence of a first regulatory protein and a second replication initiation site in the presence of a second regulatory protein, and has a stronger ability to initiate plasmid replication in the second replication initiation state than in the first replication initiation state; 2) a first regulatory protein expression cassette expressing the first regulatory protein; 3) a sequence encoding a repressor protein; 4) a screened tag gene; 5) a target gene, or a cloning site for inserting the target gene; and 6) paired recombination sites. Also provided are a host cell suitable for the recombination of the precursor plasmid and the production of a screening-tag-free plasmid, and a method for large-scale production of the screening-tag-free plasmid.

The invention belongs to the fields of biotechnology, protein engineering and plant protection science, and particularly relates to an antagonistic protein mutant for cultivating a sweet potato composite virus disease (SPVD) resistant plant and application of the antagonistic protein mutant. The key pathogenic factor of the SPVD is the RNase3 protein coded by the sweet potato chlorotic stunt virus (SPCSV), and the RNase3 protein is used as an RNA silence suppressor (RSS) to destroy host immunity. The invention provides an SPCSV-RNase3 antagonistic protein mutant, which is derived from a natural antagonistic protein, namely a sweet potato trypsin inhibitor (IbSPLTI-a). According to the present invention, a compound structure model (such as construction through AlphaFold2) of wild type IbSPLTI-a and RNase3 is analyzed, and the IbSPLTI-a is modified by using a protein directed evolution strategy, particularly by using a hotspot amino acid scanning and protein surface design strategy, such that the high-activity inhibition mutant (such as IbSPLTI-a-m0805) is successfully created and screened; in-vivo and in-vitro function verification shows that the binding affinity of the mutant (such as IbSPLTI-a-m0805) and SPCSV-RNase 3 is remarkably enhanced, and the mutant shows a virus accumulation inhibition effect superior to that of a wild type IbSPLTI-a. The invention also provides a nucleic acid sequence for coding the mutant, a plant expression vector containing the sequence, and a method for culturing an anti-SPVD transgenic plant (especially sweet potato) by using the mutant. The invention provides a core gene resource and a technical path for genetic improvement of SPVD and development of a novel antiviral protein preparation.

The present application belongs to the technical field of genetic engineering, and particularly relates to an engineered bacillus subtilis, a preparation method and application thereof. In the present application, in order to improve the supply of precursor cytidine triphosphate (CTP) in the process of fermentation of bacillus subtilis for producing cytidine, first, the feedback-inhibited CTP synthetase PyrG E156K is overexpressed to promote the conversion of uridine triphosphate to CTP; then, the repressor gene of the pyrimidine operon pyrR is knocked out to promote the synthesis of uridine monophosphate, and further promote the synthesis of CTP; finally, the pyrimidine nucleotide degradation genes pdp and cdd are knocked out to block the conversion of uridine / cytidine to uracil / cytosine and the conversion of cytidine to uridine, respectively, reduce the consumption of pyrimidine nucleotides, and further facilitate the intracellular accumulation of CTP. The yield of cytidine of the finally constructed engineering bacteria is about 12.6 times that of the original starting bacteria, and the results of the present application further prove the importance of CTP for cytidine synthesis.

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.

The present disclosure provides super-repressors capable of inhibiting the transcription of NR4A1, NR4A2, and NR4A3 target genes. The super-repressors can be used to enhance the effector functions of immune cells, e.g., for adoptive cell therapy. Methods of treating disorders using the modified immune cells are also provided.

The application discloses a stringent induction system for corynebacterium glutamicum and application thereof, and belongs to the fields of genetic engineering and microbiology. The system can be strictly regulated without an inducer and can be expressed after the inducer is added. More specifically, it relates to reducing the leakage expression of Cre recombinase by containing logic gates, repressor regulation, and RNA regulatory elements, and its use for mediating the production of large-scale genomic random rearrangement of SCRaMbLE in the synthetic genome of corynebacterium glutamicum. In particular, it is used for reducing the leakage expression level of Cre recombinase and maintaining a moderate expression level after induction. More importantly, the stringent induction system of the application can be applied to obtain rearranged strains with highly diverse genotypic sequences in genomic rearrangement, so as to achieve the purposes of improving the yield of target products or enhancing the stress resistance of strains.

The invention relates to the technical field of bioengineering, and discloses a construction method and application of recombinant Escherichia coli engineering bacteria with high yield of ergothioneine, and the method comprises the following steps: by taking Escherichia coli as a chassis, integrating a mycobacterium smegmatis egtABCDE gene cluster at an ldhA site; a repressor protein gene metJ and a cysteine lyase gene yhaM are knocked out, a methionine adenosine transferase gene metK and a serine acetyltransferase gene cysE are overexpressed by using a strong promoter, and mutant metA, thrA, serA and hisG genes for relieving feedback inhibition are introduced, so that metabolic inhibition of methionine, cysteine and histidine pathways is systematically relieved; the invention also establishes a matched fermentation process. According to the invention, endogenous precursor supply and exogenous synthesis pathways are synergistically enhanced, the precursor supply bottleneck is solved, the yield of ergothioneine is increased, and the method is suitable for industrial production.

The invention relates to the technical field of gene engineering, in particular to a phiF * expression cassette, an expression system and application of the phiF * expression cassette in xanthomonas campestris. The phiF * expression cassette is composed of a constitutive promoter Pj23119, an RBS sequence and a repressor protein gene phiFR *; the nucleotide sequence and the amino acid sequence of the repressor protein gene phiFR * are respectively shown as SEQ ID NO. 1 and SEQ ID NO. 2. According to the invention, a phiF * frame and a phiF * expression system which can be used in xanthomonas campestris are constructed, and a certain basis is provided for construction of xanthomonas campestris genetic engineering strains.

The invention provides a propionic acid engineering bacterium as well as a construction method and application thereof. The strain is obtained by taking escherichia coli E.coli W3110 as an original strain through a gene modification method: a Lac l repressor protein gene lacl, a succinate dehydrogenase gene sdhA and an isocitrate lyase inhibitory factor gene iclR are knocked out from a genome, and the escherichia coli E.coli W3110 is obtained through a gene modification method. The method comprises the following steps of: replacing a promoter of methylmalonyl-coenzyme A carboxylase scpA on a genome with a Ptrc promoter, overexpressing a pTrc99a-RM25-1795-1792 plasmid in a constructed strain, knocking out a propionate-coenzyme A synthetase gene prpE, knocking out acs and prpC genes, and carrying out gene knockout; the operation for producing propionic acid by fermentation of the strain is simple, propionic acid can be directly synthesized by using a cheap carbon source, the yield reaches 8.53 g / L after 24 hours through fermentation culture in a 500mL shake flask, and the strain has a very good industrial application prospect.

The present invention uses gene knockout technology to construct a high-yield poly (γ-glutamic acid) (γ-PGA) engineered strain of Bacillus paralicheniformis that is independent of exogenous glutamate addition. By seamlessly knocking out the carbon metabolism repressor protein A gene on the genome of Bacillus paralicheniformis 285-3, which regulates carbon metabolism, ccpA , enabling the strain to enhance carbon metabolism pathways, increase the synthesis of the substrate glutamate, and ultimately enhance the synthesis of γ-PGA. The engineered Bacillus paralicheniformis strain constructed using this technology significantly increased γ-PGA production, paving the way for the subsequent use of this engineered strain to produce γ-PGA in a manner independent of exogenous glutamate addition. Compared to the starting strain 285-3, its γ-PGA production rate is higher, effectively saving costs such as labor, energy, and equipment usage and loss, thereby further reducing the production cost of γ-PGA.