30 results about "Promoter enhancer" patented technology

The present disclosure, among other things, provides machine-learning technologies for identifying and localizing particular genomic elements (e.g., gene elements and / or regulatory elements) within nucleotide sequences, such as DNA and / or RNA sequences. In certain embodiments, similar to the manner in which image processing methods can be used to localize particular objects in images at pixel level resolution, referred to as “segmentation,” systems and methods of the present disclosure predict presence and locations of certain genomic elements within nucleotide sequences, thereby “segmenting” nucleotide sequences. Accordingly, genomic element segmentation technologies described herein may be used to generate annotations that identify and label portions of nucleotide sequences according to their predicted (e.g., via machine learning models described herein) function—e.g., as protein-coding genes, untranslated regions, splice sites, promotors, enhancers, etc. Among other things, these genomic annotations may be used to inform underlying biological processes driving diseases and facilitate development of new therapies.

The present disclosure, among other things, provides machine-learning technologies for identifying and localizing particular genomic elements (e.g., gene elements and / or regulatory elements) within nucleotide sequences, such as DNA and / or RNA sequences. In certain embodiments, similar to the manner in which image processing methods can be used to localize particular objects in images at pixel level resolution, referred to as "segmentation," systems and methods of the present disclosure predict presence and locations of certain genomic elements within nucleotide sequences, thereby "segmenting" nucleotide sequences. Accordingly, genomic element segmentation technologies described herein may be used to generate annotations that identify and label portions of nucleotide sequences according to their predicted (e.g., via machine learning models described herein) function – e.g., as protein-coding genes, untranslated regions, splice sites, promotors, enhancers, etc. Among other things, these genomic annotations may be used to inform underlying biological processes driving diseases and facilitate development of new therapies.

A method for improving the replication of covalent closed circular plasmids is provided. [Solution] The method comprises the step of preparing a covalent closed circular plasmid having a Pol I-dependent origin of replication and an insert containing a structured DNA sequence selected from the group consisting of a reverse repeat sequence, a directional repeat sequence, a homopolymer repeat sequence, a eukaryotic origin of replication, or a eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also comprises the step of modifying the covalent closed circular recombinant molecule so that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, thereby improving the replication of the covalent closed circular plasmid with the resulting Pol III-dependent origin of replication. A covalent closed circular recombinant DNA molecule without antibiotic markers is also provided.

The invention discloses a research method of a microtubule positive end TBCB and EB molecular binding mode, and relates to the technical field of neurobiology. Comprising the following steps: constructing expression vectors: respectively constructing expression vectors containing TBCB, EB1 and EB3 gene sequences; the vectors should comprise appropriate promoters, enhancers and other necessary regulatory elements to ensure efficient expression of target genes in host cells; transfecting a host cell: selecting a proper cell line as the host cell, and transfecting the constructed expression vector into the host cell independently or in a combined manner; transfection may be performed by electroporation, liposomal mediation, or other potent methods. Through detailed step division and implementation, the interaction between TBCB and EB1 / EB3 can be systematically researched, and each step aims at comprehensively revealing a binding mechanism among molecules from construction of an expression vector to final data analysis.

This invention describes a novel gene regulatory sequence containing the promoter and enhancer sequences of the human cardiac troponin T gene (TNNT2) that selectively induces expression in cardiomyocytes. This novel TNNT2 promoter / enhancer composition can be used to induce adeno-associated virus gene expression, construct cell-type-specific expression vectors, or perform cardiac-specific transgenesis. The use of this novel promoter / enhancer composition is demonstrated by the expression of mAKAP shRNA and a mAKAP-derived anchor-disrupting peptide useful for the treatment of heart failure.

The present invention relates to a recombinant alpha 1-antithricin (AAT) protein or fragment thereof expressed in genetically modified yeast for use in the treatment of viral infections and / or pulmonary inflammation. In one embodiment, the viral infection is a viral respiratory infection, preferably a coronavirus infection such as SARSCoV infection, more preferably SARS-CoV-2. The present invention further relates to a method for producing a recombinant AAT protein or fragment(s) thereof from a genetically modified yeast. In an embodiment, the method comprises culturing a genetically modified yeast comprising an exogenous nucleic acid molecule having an AAT coding region operably linked to a promoter or promoter / enhancer combination, expressing the recombinant AAT in the cultured yeast, and isolating the recombinant AAT from the culture. In an embodiment, the recombinant AAT produced from the yeast is prepared in a pharmaceutical composition, such as a solution, preferably suitable for nebulization and subsequent inhalation by a subject.

Provided herein are genetically engineered promoter-enhancer constructs for altering transcription of a gene operably linked thereto having specificity for expression in photoreceptor cells. Also provided are methods of use for these photoreceptor-specific promoter-enhancer genetic constructs; a pharmaceutical composition comprising these constructs, and methods of using the pharmaceutical composition for treating retinal diseases.

Disclosed is a recombinant adeno-associated virus (rAAV) vector that is a universal reference standard in quantitative PCR assays used to determine the genome copy titer of rAAV-transgene particles. The rAAV-Universal Standard vector includes multiple nucleotide sequences that are targets for PCR assays to support polymerization such as fragments of commonly used promoters, enhancers, polyadenylation sequences, and reporter genes found in rAAV constructs. Those multiple targets ensure that there is a positive reaction and thus provide a standard, positive control and calibrator to assure proper performance of the quantitative PCR assays without the need to use a separate reference standard for each particular rAAV particle assayed. The rAAV-Universal Standard is produced in multiple AAV serotypes and may encode a functional reporter gene to further provide a universal positive control for analytical assays of rAAV-transgene particles.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from inverted repeat sequences, direct repeat sequences, homopolymeric repeat sequences, eukaryotic origins of replication or eukaryotic promoter enhancer sequences, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.

The present application relates to a DNA sequence-based promoter-enhancer interaction prediction method, comprising: using a window to intercept the obtained multiple groups of promoter sequences and multiple groups of enhancer sequences to obtain multiple groups of promoter sequence fragments and multiple groups of enhancer sequence fragments; each group of promoter sequence fragments and each group of enhancer sequence fragments are subjected to k-mer processing to obtain multiple groups of k-mer processed promoter sequence fragments and enhancer sequence fragments; global feature extraction and multi-scale feature extraction are performed on multiple groups of k-mer processed promoter sequence fragments and enhancer sequence fragments to obtain promoter global representation, first multi-scale feature, enhancer global representation and second multi-scale feature; then the obtained multiple features are subjected to feature adaptive fusion and cross attention calculation to obtain cross attention results; based on the cross attention results, the prediction results are calculated. The method can effectively improve the model prediction accuracy.

The present invention relates to nucleic acid cassettes for gene therapy, in particular to a promoter and promoter / enhancer combination for improving transgene expression in a pulmonary parenchyma-specific / preferential manner. The invention also relates to nucleic acid cassettes comprising said promoters and promoter / enhancer combinations, viral and non-viral vectors comprising such nucleic acid cassettes, and the use of such nucleic acid cassettes and vectors for increasing the expression of therapeutic proteins by pulmonary parenchymal cells.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid, the plasmid having a Pol I-dependent replication origin and an insert, the insert comprising a structured DNA sequence selected from the group consisting of: an inverted repeat sequence, a forward repeat sequence, a homopolymeric repeat sequence, a eukaryotic replication origin, or a eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000bp from the Pol I-dependent replication origin along the replication direction. The method also includes modifying the covalently closed circular recombinant molecule so that the Pol I-dependent replication origin is replaced by the Pol III-dependent replication origin, and the Pol III-dependent replication origin covalently closed circular plasmid thus obtained has improved replication. A covalently closed circular recombinant DNA molecule without an antibiotic marker is also provided.

A method and vectors for controlling blood glucose levels in a mammal are disclosed. In one embodiment, the method comprises the steps of: treating the hepatocyte cells of a patient with a first, second or third vector, wherein the first vector comprises a promoter enhancer, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase and an albumin 3′UTR and lacks an HGH intron, wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site and an albumin 3′UTR and lacks a promoter enhancer, wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site, an albumin 3′UTR and a promoter enhancer and observing the patient's insulin levels, wherein the patient's insulin levels are controlled.

Disclosed is a recombinant adeno-associated virus (rAAV) vector that is a universal reference standard in quantitative PCR assays used to determine the genome copy titer of rAAV-transgene particles. The rAAV-Universal Standard vector includes multiple nucleotide sequences that are targets for PCR assays to support polymerization such as fragments of commonly used promoters, enhancers, polyadenylation sequences, and reporter genes found in rAAV constructs. Those multiple targets ensure that there is a positive reaction and thus provide a standard, positive control and calibrator to assure proper performance of the quantitative PCR assays without the need to use a separate reference standard for each particular rAAV particle assayed. The rAAV-Universal Standard is produced in multiple AAV serotypes and may encode a functional reporter gene to further provide a universal positive control for analytical assays of rAAV-transgene particles.

Provided herein are constructs for improved expression of transgenes (e.g., ocular transgenes), nucleic acid vectors thereof, pharmaceutical compositions thereof, and methods of use thereof (e.g., methods of treatment). The expression constructs include regulatory elements such as promoters, enhancers, insulators, and intronic sequences. Expression constructs, nucleic acid vectors and pharmaceutical compositions thereof, and methods of use thereof, disclosed herein can provide effective, durable treatments for ocular diseases.

The invention discloses a method for improving plant pilot editing efficiency, which comprises the following steps: replacing AtU6 with an NtU26 promoter through a restriction enzyme site directional cloning technology, and respectively connecting the NtU26 promoter with two serially connected PegRNA sites. The method comprises the following steps: constructing a '35S promoter-Cmylc enhancer-NtU6-26' expression unit to replace a '35S promoter-Cmylc enhancer-AtU6', and ensuring that the transcription initiation of pegRNA is regulated and controlled by a 35S promoter-Cmylc enhancer-NtU6-26 promoter; the unique transcriptional regulation ability of the NtU6-26 promoter in tobacco cells is utilized, sufficient PegRNA substrates are provided for assembly of an editing compound (nCas9-RT / pegRNA) by improving the pegRNA transcriptional level and increasing the accumulation amount of the PegRNA, the order of magnitude improvement of tobacco pilot editing efficiency is achieved, precise replacement of AtU6 by the NtU26 promoter is achieved by utilizing restriction enzyme sites, uncertainty caused by random integration is avoided, and the method has the advantages that the method is simple and convenient to operate and high in practicability. The stability and repeatability of the gene expression unit are ensured; the NtU6-26 is designed for tobacco cells, and the unique transcriptional regulation ability of the NtU6-26 can significantly improve the transcriptional level of pegRNA and increase the accumulation amount of pegRNA in the cells.

Provided herein are genetically engineered promoter-enhancer constructs for altering transcription of a gene operably linked thereto having specificity for expression in photoreceptor cells. Also provided are methods of use for these photoreceptor-specific promoter-enhancer genetic constructs; a pharmaceutical composition comprising these constructs, and methods of using the pharmaceutical composition for treating retinal diseases.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I- dependent origin of replication, and an insert including a structured DNA sequence selected from inverted repeat sequence, direct repeat sequence, homopolymeric repeat sequence, eukaryotic origin of replication or eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol Ill-dependent origin of replication, whereby the resultant Pol Ill-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided. Recombinant DNA molecules and methods related thereto which include one or more primosomal sites and / or a selectable marker oritented to transcribe away from a structured DNA sequence are also provided.

This invention provides a variety of novel adeno-associated virus (AAV) vectors for gene therapy applications in the treatment of glycogen storage disease type Ia (GSD-Ia). This document discloses numerous recombinant nucleic acid molecules, vectors, and recombinant AAVs incorporating modified G6PC promoter / enhancer sequences. When expressed from various host cell platforms, the use of said modified G6PC promoter / enhancer sequences results in enhanced AAV yield and quality. This document also provides compositions comprising the novel AAVs of this invention and methods for treating GSD-Ia using said compositions.

The invention discloses a plant pilot editing method which comprises the following steps: replacing AtU6 with an NtU26 promoter through a restriction enzyme site directional cloning technology, and respectively connecting the NtU26 promoter with two PegRNA sites which are connected in series. The method comprises the following steps: constructing a '35S promoter-Cmylc enhancer-NtU6-15 'expression unit to replace a '35S promoter-Cmylc enhancer-AtU6', and ensuring that the transcription initiation of pegRNA is regulated and controlled by a 35S promoter-Cmylc enhancer-NtU6-15 promoter; the unique transcriptional regulation ability of the NtU6-15 promoter in tobacco cells is utilized, sufficient PegRNA substrates are provided for assembly of an editing compound (nCas9-RT / pegRNA) by improving the pegRNA transcriptional level and increasing the accumulation amount of the PegRNA, the order of magnitude improvement of tobacco pilot editing efficiency is achieved, precise replacement of AtU6 by the NtU26 promoter is achieved by utilizing restriction enzyme sites, uncertainty caused by random integration is avoided, and the method has the advantages that the method is simple and convenient to operate and high in practicability. The stability and repeatability of the gene expression unit are ensured; the NtU6-15 is designed for tobacco cells, and the unique transcriptional regulation ability of the NtU6-15 can significantly improve the transcriptional level of pegRNA and increase the accumulation amount of pegRNA in the cells.

This invention provides a variety of novel adeno-associated virus (AAV) vectors for gene therapy applications in the treatment of glycogen storage disease type 1a (GSD-Ia). Disclosed herein are a number of recombinant nucleic acid molecules, vectors and recombinant AAV that incorporate a modified G6PC promoter / enhancer sequence. Utilization of the modified G6PC promoter / enhancer sequence results in enhanced AAV yield and quality when expressed from various host cell platforms. Also provided herein are compositions comprising the novel AAV of the invention and methods of treating GSD-Ia using the same.

A recombinant alphal-antitrysin (AAT) protein or a fragment thereof expressed in a genetically modified yeast for use in the treatment of a viral infection and / or lung inflammation. In one embodiment, the viral infection is a viral respiratory infection, preferably a coronavirus infection, such as a SARS CoV infection, more preferably SARS-CoV-2. Also disclosed is a method of producing recombinant AAT protein or fragment(s) thereof from genetically modified yeast. In embodiments, the method includes the steps of culturing a genetically modified yeast comprising an exogenous nucleic acid molecule with an AAT-encoding region operably linked to a promoter or promoter / enhancer combination, expressing recombinant AAT in the cultured yeast, and isolating recombinant AAT from the culture. In embodiments, the recombinant AAT produced from yeast is prepared in a pharmaceutical composition, such as a solution, preferably suitable for nebulization and subsequent inhalation by a subject.

Provided is a gene drug for treating retinal degenerative diseases. Provided is a gene expression cassette, which comprises a coding sequence of a photosensitive protein, and optionally, a promoter, an enhancer, and / or an intron. Optionally, the photosensitive protein comprises a microbial photosensitive protein or an animal photosensitive protein. The provided gene drug has demonstrated a good therapeutic effect in cells and mice, making the optogenetic treatment method targeting bipolar cells a potential treatment method with broad clinical application prospects.

The invention discloses a baculovirus expression vector capable of efficiently expressing swine foot-and-mouth disease virus-like particles as well as a construction method and application of the baculovirus expression vector. The baculovirus expression vector contains a segment of skeleton sequence and a segment of gene synthesis sequence of a pFastbacic-dual vector. The gene synthesis sequence comprises a first terminator, a first cloning site, an internal ribosome entry site, a transcriptional activation factor, a first promoter, an enhancer, a second promoter, a second cloning site and a second terminator which are connected in sequence, the first cloning site and the second cloning site are introduced into a gene sequence of a pig foot-and-mouth disease 3C protein and a gene sequence of a pig foot-and-mouth disease P12A protein respectively. After the baculovirus prepared by the baculovirus expression vector provided by the invention infects insect cells, the expression quantity of 3C protein can be reduced, the expression quantity of P12A protein can be increased, the expressed 3C protein can cut the P12A protein to further form swine foot-and-mouth disease virus-like particles, and the expression quantity of the foot-and-mouth disease virus-like particles can be remarkably increased.

The present disclosure, among other things, provides machine-learning technologies for identifying and localizing particular genomic elements (e.g., gene elements and / or regulatory elements) within nucleotide sequences, such as DNA and / or RNA sequences. In certain embodiments, similar to the manner in which image processing methods can be used to localize particular objects in images at pixel level resolution, referred to as "segmentation," systems and methods of the present disclosure predict presence and locations of certain genomic elements within nucleotide sequences, thereby "segmenting" nucleotide sequences. Accordingly, genomic element segmentation technologies described herein may be used to generate annotations that identify and label portions of nucleotide sequences according to their predicted (e.g., via machine learning models described herein) function – e.g., as protein-coding genes, untranslated regions, splice sites, promotors, enhancers, etc. Among other things, these genomic annotations may be used to inform underlying biological processes driving diseases and facilitate development of new therapies.

To realize a novel promoter enhancer for filamentous fungi. [Solution] An enhancer for a filamentous fungus promoter containing a polynucleotide having the base sequence shown in sequence A below: Sequence A: TXXXXCCAC; (wherein sequence A, X represents any base).

A therapy system for genomic reprogramming and DNA repair, including: a gene identification module configured to detect and isolate regenerative genes from planarian neoblasts, such as Smedwi-1, Smedwi-2 and β-catenin, which are responsible for cell regeneration and pluripotency; a gene adaptation unit configured to perform codon optimization and regulatory sequence modification of isolated regenerative genes for compatibility with human DNA, including the incorporation of human-specific promoters, enhancers, and terminators; a selective nucleopathy module configured with CRISPR-Cas9-based mechanisms and custom-designed guide RNAs, whereby the module selectively identifies, targets and removes mutated or damaged DNA in target cells while preserving healthy DNA; a DNA delivery unit configured to transfer the adapted regenerative genes into human stem cells, utilizing gene transfer techniques such as viral vectors, electroporation, or lipid-based nanoparticles for efficient gene integration; a genetic switch control system configured with programmable genetic kill switches for the selective activation and deactivation of regenerative genes, whereby the system enables controlled gene expression to prevent unwanted cell proliferation and tumor formation; a biopolymer scaffold support configured to provide a structural scaffold at the target site to facilitate the delivery of modified stem cells or regenerative genes and to promote cell growth and tissue integration; and an interface for hyperbaric oxygen therapy that increases cell metabolism, improves the accuracy of DNA repair and promotes tissue regeneration in synergy with regeneration and DNA repair processes.

The invention belongs to the field of biological medicine, and relates to a potential drug system for gene therapy of Alzheimer's disease (AD), which comprises cDNA of hHEBP1 and a carrier system thereof. The vector system is a recombinant system of plasmid and virus AAV2, and comprises an hCD68 promoter, a CMV enhancer and a WPRE sequence. When AD treatment is carried out, AAV virus particles are assembled after in-vitro culture, and core elements contained in the virus particles sequentially comprise an MV enhancer, an hCD68 promoter, cDNA of hHEBP1 and a WPRE structure. In a preclinical animal experiment, the system can realize long-term stable expression of HEBP1 in microglia cells in a hippocampus region, realizes repair of neuronal cells, and provides a new selection scheme for AD gene therapy.

The present invention describes a novel gene regulatory sequence comprising the promoter and enhancer sequences of the human cardiac troponin T gene (TNNT2) that guide selective expression in cardiac muscle cells. The novel TNNT2 promoter / enhancer composition can be used for guiding adeno-associated virus gene expression, or used for constructing a cell type specific expression vector, or used for heart specific transgenosis. Uses of the novel promoter / enhancer compositions are demonstrated by expression of mAKAP shRNA, as well as anchor disrupting peptides derived from mAKAP, useful for the treatment of heart failure.

The invention relates to the field of plant genetic engineering, and particularly discloses an agrobacterium tumefaciens-mediated NtU6-27 driving type tobacco pilot editing transformation identification method. Wherein the nCas9-RT fusion protein expression unit is driven by a 35S promoter, and the pegRNA expression unit is driven by a 35S promoter-Cmylc enhancer-NtU6-27 promoter. The preparation method comprises the following steps: preparing a tobacco explant, constructing a recombinant expression vector, introducing the vector into agrobacterium tumefaciens, carrying out enlarged culture, carrying out agrobacterium tumefaciens-mediated transformation on the tobacco explant, carrying out screening culture to obtain a regenerated plant, and identifying an editing effect through PCR (Polymerase Chain Reaction) and sequencing. The composition can be used for tobacco precise genetic improvement and gene function research, and has the advantages that pegRNA is driven by NtU6-27 to efficiently express, and the editing efficiency is high; in addition, the preparation method has the advantages of being clear in step, high in repeatability and accurate in identification.