50 results about "Repressor" patented technology

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.

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 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.

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 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 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 present invention relates to the therapeutic use of exosome comprising super-repressor-IκB (srIκB) for liver disease. The present invention provides a pharmaceutical composition for preventing or treating liver disease, comprising an exosome containing super-repressor-IκB (srIκB) as an active ingredient, wherein the liver disease is liver fibrosis, liver cirrhosis, fatty liver, alcoholic liver disease, cholestasis, or a combination thereof.

Provided herein is a cell comprising: (a) a polynucleotide encoding a binding-triggered transcriptional switch (BTTS) comprising an intracellular domain comprising a DNA binding domain (DBD) and one or more epigenetic editors and (b) a nucleic acid comprising a binding site for the DBD, a promoter and, operably linked to the promoter, a coding sequence encoding a payload. Binding of the BTTS to an antigen results in decreased expression of the payload via epigenetic modifications. If the payload is a transcriptional repressor, then binding of the BTTS results in increased expression of a second payload that is operably linked to the first payload. These changes result in long-term epigenetic changes in expression of the payloads and can be used to steer cell fate or function changes in response to BTTS detected inputs.

The invention relates to the technical field of gene engineering, and discloses a rice stripe mosaic virus (RSMV) expression vector, a gene silencing vector, a preparation method and application. The rice stripe mosaic virus vector composition provided by the invention comprises a construction body used for transcriptional generation of virus anti-genome RNA, a construction body used for expressing trans-acting protein necessary for virus replication and transcription, and a construction body used for expressing a virus silencing suppressor. By utilizing the combination of the components, the recombinant RSMV with infection activity can be obtained in nicotiana benthamiana cells, and the recombinant virus can be further introduced into a mediator insect electro-optic leafhopper through micro-injection and is inoculated to a rice plant through the electro-optic leafhopper to establish systemic infection. The invention provides application of the recombinant virus vector in expression of foreign proteins or silence of endogenous genes in electro-optic leafhoppers and rice bodies and the like. The vector has the advantages of strong exogenous fragment bearing capacity, high genetic stability and the like.

The present disclosure relates to methods for increasing DNA methylation of a site in a genomic region comprising a target gene, e.g., using an expression repressor comprising a DNA targeting moiety that binds to a target sequence in the region and an effector domain (e.g., a DNA methyltransferase) that methylates DNA, or a nucleic acid encoding the expression repressor. Also disclosed are systems comprising two or more expression repressors. Compositions may be used, for example, to reduce expression of the target gene in a cell or subject, for example, to treat a condition associated with expression of the target gene.

The disclosure features compositions, systems, and uses thereof, comprising polynucleotide(s) encoding target molecule(s), and polynucleotide(s) encoding repressor(s) or timer(s). The repressor(s) or timer(s) delay expression of the target molecule(s).

The invention discloses a method for improving activity of aspergillus oryzae amylase through CreA C terminal dephosphorylation mutation, and belongs to the technical field of enzyme engineering. According to the method, a CreA dephosphorylation mutant strain is constructed by performing site-specific mutagenesis on a C-terminal phosphorylation site coding gene of an aspergillus oryzae carbon metabolism inhibition core transcription factor CreA, so that a carbon metabolite repression effect is relieved, and the yield of amylase is increased. Specifically, four phosphorylation sites (S414, S415, T416 and S419) at the C tail end of the CreA protein are mutated into valine (V). When the mutant strain is used for fermentation, the expression of amylase can be remarkably improved. Experiments prove that the amylase activity of the mutant strain 4M can be 3 times that of a reference strain after the mutant strain is cultured for 6 days in a culture medium taking starch as a carbon source. The invention provides a new strain modification strategy for efficient production of amylase.

This invention discloses a method for synergistic improvement of wheat quality and disease resistance based on the creation and utilization of the T1DL.1VS translocation line. Addressing the problem of low breeding efficiency caused by the dispersion of disease resistance and quality genes, this invention creates a novel T1DL.1VS translocation line that aggregates Pm67 / Glu-V1 by crossing the T1DL.1V#4S translocation line NAU195 carrying Glu-V1 with the T1DL.1V#5S translocation line NAU196 carrying Pm67. Recombinant single plants are screened in the F2 generation. Furthermore, utilizing the co-segregation characteristic of this translocation line with the wax repressor gene IW-V1, a dual-marker selection system combining morphological markers and the NAU1VS-1 molecular marker is established and introduced into a high-yielding variety background. This translocation line has no significant negative effect on yield traits, and the newly bred lines exhibit high resistance to powdery mildew and significantly improved bread processing quality, achieving synergistic and efficient improvement of wheat disease resistance and processing quality.

This specification provides a gene repressor system comprising a fusion protein, such as a fusion protein containing a non-catalyzed class 2, type V CRISPR protein useful for repressing the precursor protein-converting enzyme subtilisin / kexin type 9 (PCSK9) gene, and a guide nucleic acid (gRNA). Methods for constructing and using such a system for repressing PCSK9 transcription are also provided.

Provided herein are gene repressor systems comprising repressor fusion proteins, such as DNA-binding proteins and, in some cases, non-catalyzed CRISPR proteins and guide nucleic acids (gRNAs), which are useful in repressing genes. Also provided are methods for using such systems to repress gene transcription.

The invention provides a gene editing system of xanthomonas campestris and application of the gene editing system, and relates to the technical field of gene engineering. The gene editing system comprises a Cpf1 protein expression vector and a crRNA expression vector, the Cpf1 protein expression vector comprises an Fncpf1 expression cassette and a lambda Red expression cassette, the Fncpf1 expression cassette is composed of a sequential promoter, an RBS sequence and a Cpf1 protein coding gene, and the lambda Red expression cassette is composed of a repressor protein AraC, a promoter Para and a lambda Red coding gene. By using the system, the gene knockout efficiency of xanthomonas campestris reaches 70%, a related gene xanK synthesized by mycoflavin is successfully knocked out, an efficient production strain of colorless xanthan gum is obtained, the modification requirements of multiple fields such as food, medicines and cosmetics on xanthomonas campestris are met, and the system has an extremely high application prospect.

This invention relates to a recombinant photosynthetic bacterial strain with high tocopherol production, its construction method, and its applications. The recombinant strain knocks out the photosynthetic gene repressor transcription factor PpsR and integrates the mevalonate MEV pathway gene from *Paragonimococcus zeatans*, the p-hydroxyphenylpyruvate dioxygenase (HPPD) gene from *Pseudomonas putida*, the urokinase chlorophyllase (HPT) gene from *Synostemma pentaphyllum*, the tocopherol cyclase (TC) gene from *Arabidopsis thaliana*, and feedback-resistant 3-deoxy-D-arabinohepenoyl-7-phosphate synthase (AroG*). D146N Genes and prephenyl acid dehydrogenase TyrA* M53I Genes. This recombinant photosynthetic bacterium strain achieves highly efficient heterologous synthesis of tocopherol by knocking out regulatory genes and integrating multiple metabolic modules, with a fermentation yield of up to 3.51 g / L, showing broad prospects for industrial application.

This invention belongs to the fields of bioinformatics and protein function identification, specifically relating to a machine learning-based method for identifying viral RNA silencing repressors. The method includes: obtaining the amino acid sequence of the viral protein to be tested, inputting it into a pre-trained ESM-2 model to extract deep feature vectors, then inputting the feature vectors into a trained XGBoost classifier, and outputting an identification result indicating whether the viral protein is a viral RNA silencing repressor. This invention integrates the automatic feature extraction capability of ESM-2 with the efficient classification performance of XGBoost, significantly improving prediction accuracy and generalization ability. It is mainly used in fields such as plant antiviral genetic engineering, disease-resistant breeding, and biopesticide development, enabling rapid screening of potential viral RNA silencing repressors and providing an efficient tool for functional gene mining.

Enhancing fetal hemoglobin (HbF) production through targeted promoter editing using base editing technologies. An embodiment includes a composition for enhancing fetal hemoglobin (HbF) expression in mammalian cells, the composition comprising a base editing system comprising at least two guide RNAs comprising targeting sequences selected from SEQ ID NO: 1 to SEQ ID NO: 5. Specifically, it involves the design and use of single guide RNAs (sgRNAs) targeting key regulatory elements within the gamma-globin (HBG1 / 2) promoter. The method combines the use of adenosine base editors to introduce mutations, creating new binding sites for transcriptional activators like TAL1 and KLF1, and disrupting repressor binding sites such as BCL11A and ZBTB7A / LRF. This dual-targeting approach significantly increases HbF expression, offering a potential therapeutic strategy for treating beta-hemoglobinopathies like sickle cell disease and beta-thalassemia.

Provided herein are gene repressor systems comprising fusion proteins, such as fusion proteins comprising a DNA binding domain such as a TALE, zinc finger or catalytically-dead CRISPR protein and guide nucleic acid (gRNA), which are useful in the repression of a proprotein convertase subtilisin / kexin Type 9 (PCSK9) gene. Also provided are methods of using such systems to repress transcription of PCSK9.

The invention discloses a recombinant genetically engineered bacterium for stably producing beta-alanine without antibiotics and inducers and application of the recombinant genetically engineered bacterium, the recombinant genetically engineered bacterium takes Escherichia coli probiotics E15 as a starting strain, and one or more of the following gene editions are carried out: (1) weakening an icd gene; (2) introducing a toxin-antitoxin system hok / sok; (3) knocking out the lacI gene; and (4) overexpression of a temperature-sensitive regulation and control system: the temperature-sensitive regulation and control system is obtained by replacing a lacI gene promoter with a PR promoter and introducing a CI857 repressor protein coding gene expressed by a PL promoter. Finally, the strain constructed by the invention can realize stable and efficient production of beta-alanine in the fermentation process without adding antibiotics and inducers.

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.

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.