42 results about "Transcriptional 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 invention belongs to the field of biosynthesis, and in particular relates to LBD (Levandanenediol Pyrophosphate) synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LSLPPs mutant containing at least one mutation of D730L, D374L, N674L or G379M is obtained by performing mutation and optimization on LSLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 26.11 g / L and is greatly improved compared with the prior art.

The invention belongs to the field of biosynthesis, and particularly relates to sclareol synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, sclareol biosynthesis yield is generally low, sclareol synthase is mutated and optimized, an SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 21.13 g / L and is greatly improved compared with the prior art.

The invention relates to a gene expression cassette and application thereof. The gene expression cassette comprises a tyrosinase gene, a transcriptional repressor protein gene, a melanin response promoter and a reporter gene, and the reporter gene is located at the downstream of the melanin response promoter and is regulated by the promoter. The gene expression cassette with a specific structure is designed, melanin in cells can be responded, a melanin biosensor can be further constructed by utilizing the gene expression cassette, the gene expression cassette has good specificity, and high-throughput screening of strains can be realized. Furthermore, ALE evolution is carried out on the strain for producing melanin, high-throughput screening is carried out based on a designed sensor, the strain for producing melanin with high yield is obtained, through shake-flask culture and 5L fermentation tank culture, the yield of melanin reaches up to 10.84 g / L and 29.04 g / L respectively, and an efficient and feasible solution is provided for large-scale production of melanin.

The present invention discloses a recombinant saccharomyces cerevisiae for efficiently biosynthesizing inositol and its construction method and application, and belongs to the field of microbial genetic engineering. The recombinant saccharomyces cerevisiae of the present invention strengthens the expression of hexokinase gene hxk, inositol 3-phosphate synthase gene ino, inositol monophosphatase gene suhb, and realizes the rearrangement of glucose metabolism by knocking out the transcriptional repressor opi1 of inositol 3-phosphate synthase (Ino1) and glucose 6-phosphate dehydrogenase gene zwf1, and relieves feedback inhibition; In addition, the present invention further increases the yield of inositol by screening inositol external emission protein gene ompf, ompc and knocking out inositol internal transport protein gene itr1, itr2. The present invention realizes the efficient production of inositol (yield is as high as 195.7g / L), with good industrialization prospect.

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.

Recombinant microorganism in which a gene encoding a transcriptional repressor of the glycine cleavage system, pyruvate formate lyase or phosphoglycerate dehydrogenase, is attenuated or deleted from a host microorganism with a formic acid assimilation pathway, a ppsR gene encoding a phosphoenolpyruvate synthase regulatory protein, or a purT gene encoding a phosphoribosylglycinamide formyltransferase, is attenuated or deleted, a gcvTHP gene, consisting of the gcvT gene, the gcvH gene and the gcvP gene, which encodes an enzyme involved in a glycine cleavage system reaction in which host microorganism with the formic acid assimilation pathway is highly expressed, and a gene encoding formate tetrahydrofolate ligase, methenyl tetrahydrofolate cyclohydrolase or methylene tetrahydrofolate dehydrogenase is introduced into the host microorganism via the formic acid assimilation pathway.

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', wherein the recruitment domain A and the recruitment domain A' are capable of interacting with each other such that one of the first fusion and the second fusion, or a portion thereof, can be recruited to the vicinity of the other fusion. Further provided is the use of the complex in the manufacture of a product for silencing expression of a target gene and in the manufacture of a medicament.

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.

Method of increasing protein content in a eukaryotic cell comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site; method of producing a plant with increased protein content comprising crossing and selecting for increased protein content; method of increasing resistance to a pathogen or a pest in a plant cell or plant comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site, alone or in further combination with expressing QQS in the plant cell or plant; method for producing a plant with increased resistance to a pathogen or a pest comprising crossing and selecting for increased resistance to the pathogen or the pest; a cell, collection of cells, tissue, organ, or organism, such as a plant, in which the NF-YC4 gene comprises a promoter comprising a transcriptional repressor binding site that has been modified so that the transcriptional repressor cannot prevent transcription of the NF-YC4; plants and hybrids thereof; and seeds.

This invention belongs to the field of plant molecular biology and genetic breeding technology, specifically relating to a molecular marker related to the malic acid content of apple fruit and its application. The molecular marker is... MdNAC029 The single nucleotide polymorphism site in the gene promoter region, this site is located MdNAC029 The nucleotide sequence of the gene promoter region, starting from the 2693rd base at the 5' end, corresponds to... MdNAC029 At a position 442 bp upstream of the transcription start codon, the polymorphism is either T or C, with the C / C genotype indicating high malic acid content and the T / T genotype indicating low malic acid content. This invention, through molecular biology experiments, confirms that the T allele is the binding site for the transcriptional repressor MdMYC2, and the C allele eliminates the binding of MdMYC2, thereby relieving transcriptional repression and promoting malic acid accumulation. This provides a new molecular marker for the genetic improvement of apple fruit acidity traits, accelerating the breeding process of superior varieties.

Method of increasing protein content in a eukaryotic cell comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site; method of producing a plant with increased protein content comprising crossing and selecting for increased protein content; method of increasing resistance to a pathogen or a pest in a plant cell or plant comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site, alone or in further combination with expressing QQS in the plant cell or plant; method for producing a plant with increased resistance to a pathogen or a pest comprising crossing and selecting for increased resistance to the pathogen or the pest; a cell, collection of cells, tissue, organ, or organism, such as a plant, in which the NF-YC4 gene comprises a promoter comprising a transcriptional repressor binding site that has been modified so that the transcriptional repressor cannot prevent transcription of the NF-YC4; plants and hybrids thereof, and seeds.

The invention discloses a method for enhancing cholesterol uptake of saccharomyces cerevisiae, and belongs to the technical field of bioengineering. The method comprises the following steps: knocking out transcription inhibition factors MOT3 and ROX1 of genes related to cholesterol uptake and transport of saccharomyces cerevisiae; the transcription activating factors SUT1 and UPC2-1 of the cholesterol transporter gene are over-expressed; expression of sterol esterifying enzyme ARE2 is enhanced, so that uptake and storage of exogenous cholesterol by the saccharomyces cerevisiae are further enhanced. According to the method disclosed by the invention, the uptake and storage of cholesterol by the saccharomyces cerevisiae are enhanced, and an important reference is provided for construction of a steroid hormone synthesis chassis taking cholesterol as a substrate.

This invention belongs to the field of biosynthesis, specifically relating to a variant of lysine pyrophosphate diol synthase and the constructed engineered yeast strain for perillaldehyde synthesis. To address the generally low yield of perillaldehyde biosynthesis in existing technologies, this invention mutates and optimizes the lysine pyrophosphate diol synthase to obtain an SsLPPs mutant containing the G379M mutation, which is then used for perillaldehyde synthesis. Simultaneously, to address the genetic instability issues associated with free plasmid expression in some Saccharomyces cerevisiae strains, this invention constructs a perillaldehyde synthesis pathway within the yeast genome. Furthermore, by enhancing the acetyl-CoA synthesis pathway, the MVA pathway, and the perillaldehyde synthesis pathway, and by knocking out some transcriptional repressors, the substrate strain synthesizing perillaldehyde is modified, resulting in a significantly increased yield compared to existing technologies.

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 present application belongs to the technical field of bioengineering, and particularly relates to an engineered bacterium for producing cytidine diphosphate choline as well as a construction method and application thereof. The present application takes Bacillus subtilis BSC1-4 as a starting bacterium, blocks the conversion of choline to glycine betaine to reduce unnecessary loss of substrates in the fermentation process, improves the absorption of choline, and enables the substrates to continuously generate cytidine diphosphate choline in the direction of generating cytidine diphosphate choline. The construction method of the engineered bacterium is specifically as follows: the key genes of the choline degradation pathway in the genome of Bacillus subtilis are knocked out gbsA and gbsB , the transporter protein OpuD is overexpressed, and finally the transcriptional repressor genes opcR and gbsR are knocked out. Experimental results show that the engineered bacterium constructed in the present application has a cytidine diphosphate choline yield of 1181.6±11.5 mg / L after 24 h of fermentation in a shake flask.

The invention belongs to the field of biosynthesis, and in particular relates to an improved variant of libdanenediol pyrophosphate synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LsLPPs mutant containing N674L mutation is obtained by performing mutation and optimization on LsLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; and the chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, so that the yield is greatly improved compared with the prior art.

The invention provides application of a transcription inhibition factor PuZAT68 gene in regulation and control of drought resistance of populus japonica, and belongs to the technical field of gene engineering and plant breeding. The nucleotide sequence of the transcription inhibition factor PuZAT68 gene is shown as SEQ ID NO.3, and the amino acid sequence of the transcription inhibition factor PuZAT68 gene is shown as SEQ ID NO.4. It is found that a PuZAT68 inhibition expression plant shows the growth state obviously superior to that of wild type WT; the PuZAT68 overexpression plant shows an obvious growth inhibition phenomenon, physiological index determination of different plants finds that inhibition of PuZAT68 expression can reduce production of reactive oxygen species (ROS), damage of oxidative stress to cell membranes can be effectively relieved, more osmotic regulation substances are accumulated, the osmotic regulation capability of cytoplasm is remarkably enhanced, and the osmotic regulation effect is improved. The plant growth regulator plays a better role in drought resistance, so that the plant can still maintain relatively normal growth under drought stress.

The present application belongs to the field of biosynthesis, and particularly relates to a sclareol synthase and a yeast engineering bacterium for producing sclareol. In order to solve the problem of low yield of sclareol biosynthesis in the prior art, the sclareol synthase is mutated and optimized in the present application, a SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability existing in the expression of free plasmid in some Saccharomyces cerevisiae strains, the present application constructs a sclareol synthesis pathway in the yeast genome, strengthens the acetyl coenzyme A synthesis pathway, the MVA pathway and the sclareol synthesis pathway, and knocks out part of the transcriptional repressor to modify the chassis bacterium for synthesizing sclareol. The yield in a 5 L fermenter reaches 21.13 g / L g / L, which is greatly improved compared with the prior art.

This invention discloses a MYB transcriptional repressor, AsMYB054, related to the synthesis of 2-(2-phenylethyl) chromones in Aquilaria sinensis and its applications. Targeting the weak links in the synthesis and regulation of 2-(2-phenylethyl) chromone compounds, this invention clones for the first time the AsMYB054 gene, a transcriptional repressor related to the regulation of 2-(2-phenylethyl) chromone synthesis in Aquilaria sinensis. AsMYB054 is located in the cell nucleus and possesses transcriptional repressive activity, capable of binding to and inhibiting the activity of the promoters of enzyme genes AsPKS02 and AsPKS09, which are related to the biosynthesis of 2-(2-phenylethyl) chromone compounds. Inhibiting the expression of AsMYB054 using gene editing or RNA interference techniques can promote the accumulation of 2-(2-phenylethyl) chromone compounds in Aquilaria sinensis, showing broad application prospects and significant economic value.

The invention discloses application of a GBI gene in regulation and control of rice plant type development. The applicant performs over-expression on the member GBI of the rice class II LBD gene family, and the obtained over-expression transgenic plant shows a semi-dwarf phenotype. Gene basic feature analysis finds that GBI is a transcription inhibition factor positioned in a cell nucleus. According to preliminary analysis of a molecular mechanism, transcription of GBI is inhibited by BR and GA at the same time, and GBI interacts with TUD1 participating in a BR signal transduction pathway and SLR1 participating in a GA signal transduction pathway at the same time. The results show that GBI is combined with GA and BR signal transduction pathways to jointly regulate and control the development of the rice plant height.

The invention discloses a DNA (Deoxyribose Nucleic Acid) binding transcription inhibition factor LacI mutant and construction of a biosensor based on the mutant, and belongs to the technical field of synthetic biology and metabolic engineering. The biosensor is a novel biosensor responding to lactose, 2 '-fucosyllactose, 3-fucosyllactose and other analogues, the biosensor is constructed on the basis of a new sensor original LacI mutant obtained in the invention, and the LacI mutant comprises I79H, V150A, S193G, Q291I, Q291V and Q291F. The mutants have strong binding force with a promoter in the absence of lactose and analogues thereof so as to inhibit transcription, and in the presence of a substance to be detected, the mutants sensitively respond and are separated from the promoter, and reporter gene starts expression so as to realize detection. Through verification, the biosensor constructed by the invention has relatively high detection sensitivity and specificity.

The present application belongs to the field of biosynthesis, and particularly relates to pyrophosphate lyddane diol ester synthase and yeast engineering bacteria for producing sclareol. In order to solve the problem of generally low yield of sclareol biosynthesis in the prior art, the pyrophosphate lyddane diol ester synthase is mutated and optimized in the present application, and a SsLPPs mutant containing at least one mutation of D730L, D374L, N674L or G379M is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability existing in the expression of free plasmid in part of saccharomyces cerevisiae strains, the present application constructs a sclareol synthesis pathway in the yeast genome, and at the same time, the acetyl coenzyme A synthesis pathway, the MVA pathway and the sclareol synthesis pathway are strengthened, and part of the transcriptional repressor is knocked out, so that the bottom bacteria for synthesizing sclareol are modified. The yield in a 5 L fermenter reaches 26.11 g / L, and the yield is greatly improved compared with the prior art.

The present application relates to the field of biotechnology, in particular to a high-yield high-performance cellulase Trichoderma reesei engineering strain and a construction method and application thereof, the Trichoderma reesei engineering strain co-overexpresses beta-glucosidase gene Aabgl1 and expansin gene Trswo1, and knocks out transcriptional repressor gene Trace1, the Trichoderma reesei engineering strain has high cellulase yield, which is 100.15% higher than that of the starting strain Trichoderma reesei Rut C30.The high-performance cellulase produced by the Trichoderma reesei engineering strain has balanced enzyme system composition and excellent hydrolysis performance, when corn straw is hydrolyzed, the glucan conversion rate reaches 94.89%, the hydrolysis performance is equivalent to that of commercial enzyme preparation Cellic CTec2.0, and good industrial development prospect and practical application value are shown.

The present invention relates to recombinant poxviruses comprising a binding sequence for a transcriptional repressor protein, such as a tetracycline repressor protein, located between a poxvirus promoter and the open reading frame (ORF) of a yield-reducing transgene, and to transgenic poxvirus producer cells expressing the corresponding repressor protein.

The present application relates to an engineered gene transcriptional repression tool targeting inhibin subunit βE (INHBE) and the use thereof, and specifically relates to a complex comprising a first fusion and a second fusion, wherein: 1) 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 2) the first fusion or the second fusion comprises a nucleic acid-binding domain. Moreover, the recruitment domain A and the recruitment domain A' are capable of interacting with each other, such that one of the first fusion and the second fusion or a part thereof can be recruited to the vicinity of the other fusion; and the nucleic acid-binding domain is capable of specifically binding to an INHBE gene transcriptional regulatory element.

This invention belongs to the field of biosynthesis, specifically relating to a variant of lysine pyrophosphate diol ester synthase and engineered bacteria for producing perillaldehyde. To address the generally low yield of perillaldehyde biosynthesis in existing technologies, this invention mutates and optimizes lysine pyrophosphate diol ester synthase, obtaining an SsLPPs mutant containing the D374L mutation, and uses this mutant for perillaldehyde synthesis. Simultaneously, to address the genetic instability issues associated with free plasmid expression in some Saccharomyces cerevisiae strains, this invention constructs a perillaldehyde synthesis pathway in the yeast genome. Furthermore, it modifies the substrate bacteria synthesizing perillaldehyde by enhancing the acetyl-CoA synthesis pathway, the MVA pathway, and the perillaldehyde synthesis pathway, and by knocking out some transcriptional repressors, resulting in a significant increase in yield compared to existing technologies.

Disclosed in the present invention are a xylose-inducible transcription factor component and a use thereof. The xylose-inducible transcription factor component comprises: a xylose-inducible transcription factor XlnR derived from a filamentous fungus and a binding consensus sequence of the xylose-inducible transcription factor XlnR. By combining the transcription factor XlnR with a prokaryotic xylose-inducible transcriptional repressor XylR, a dual-regulated inducible system obtained exhibits an extremely low leakage level, but maintains a maximum activation level comparable to that of an XlnR single-regulated xylose-inducible system. The present invention solves the problems of low induction strength and slow response in transcription factor-dependent xylose transcriptional induction systems in the field of synthetic biology.

The invention provides a transcription suppressor and application thereof, and finds that after the suppressor is knocked out, the expression quantity of a related target gene (LOCOs08g06320) in rice is obviously increased, which indicates that after the suppressor is knocked out, the expression of the target gene is relieved, the element is verified to have definite inhibitory activity on the transcriptional level, and in addition, the transcription suppressor can be used for preparing the rice transcription suppressor. The invention further provides a method for transferring the suppressor into a vector to artificially regulate and control the expression of related genes for the first time. The discovery provides a new research direction for crop breeding, and has important breeding significance and development prospects. Compared with the prior art, a gene with transcriptional inhibition activity is identified and verified for the first time, the inhibitor can be applied to yield regulation and stress response in follow-up research by utilizing the inhibition characteristic of the inhibitor, and the inhibitor can play an important role in the technical fields of follow-up molecular biology and crop genetic improvement. The obvious biological and application values are realized.

The invention discloses saccharomyces cerevisiae engineering bacteria for efficiently synthesizing mogroside V based on peroxisome subcellular organelles and a construction method of the saccharomyces cerevisiae engineering bacteria. A construction method of the engineering bacterium comprises the following steps: (1) integrating mogrol metabolic enzyme and leucine selection marker Leu2 at a site of a transcription inhibition factor GAL80 of saccharomyces cerevisiae; and (2) integrating peroxisome membrane protein Pex11, carnitine acetyltransferase Cat2, phosphoglucose mutase PGM1, alpha-phosphoglucose mutase PGM2, uridine diphosphate glucose pyrophosphorylase UGP1, glycosyl transferase UGTMG1, glycosyl transferase SgUGT94-289-3 and a histidine selection marker His3 at the site of the saccharomyces cerevisiae glucoside hydrolase Exg1. The engineering strain has the advantages of high yield, strong metabolic flux directionality, wide industrial application prospect and the like, and a new technical route is provided for microbial manufacturing of natural sweeteners.