10 results about "Promoter enhancer" patented technology

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.

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.

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.

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.

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.

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.

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).

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 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.