26 results about "Polyadenylation" patented technology

The invention discloses a polyadenylation signal mutation harmfulness prediction method fusing an improved TabPFN network and SHAP interpretability. The polyadenylation signal mutation harmfulness prediction method comprises the following steps: 1, collecting a human polyadenylation signal mutation sample and carrying out pretreatment; 2, carrying out feature annotation on the mutation sample by using a computational biological tool; 3, selecting features by using a feature selection framework; 4, constructing a mutation prediction model based on the improved TabPFN; and 5, quantifying the contribution degree of each feature to a model prediction result by using an SHAP interpretability analysis technology so as to understand the output of a complex model and realize the interpretability of the model. The method provided by the invention solves the problems of complex parameter tuning, weak interpretability and the like in the prior art, so that the harmfulness of polyadenylation signal mutation is accurately, efficiently and interpretably predicted.

The application discloses a method and device for identifying a selective polyadenylation cell subpopulation, an equipment and a readable storage medium, and relates to the technical field of cell subpopulation identification. The method comprises the following steps: obtaining a selective polyadenylation expression count matrix containing a plurality of spots in spatial transcriptome data, spatial coordinate information of each spot, and a phenotype label of each spot; constructing a spatial adjacency graph of each spot based on the spatial coordinate information; training a graph attention network model by taking the selective polyadenylation expression count matrix and the spatial adjacency graph as input and taking the phenotype label as a supervision signal until the graph attention network model converges; predicting the confidence of each spot based on the graph attention network model; and taking each spot with a confidence higher than a preset confidence threshold as the selective polyadenylation cell subpopulation. The method increases the identification accuracy of the selective polyadenylation cell subpopulation.

To provide adeno-associated variants, formulations and methods for pulmonary delivery.SOLUTION: The present disclosure provides variant AAV capsid proteins that confer tropism for lung cells, as well as recombinant adeno-associated viruses comprising the variant AAVs and pharmaceutical compositions comprising the same, and their use in the delivery of heterologous nucleic acids to lung cells for the treatment of lung disorders. (ii) nucleic acids comprising, from 5 ' to 3 ', (a) AAV2 terminal repeats, (b) a promoter, (c) a nucleotide sequence encoding a human cystic fibrosis transmembrane conductance regulator (CFTR) or a biologically active truncated CFTR lacking amino acids 708 to 759 of the human CFTR sequence, (d) a poly-adenylation sequence, and (e) AAV2 terminal repeats.SELECTED DRAWING: None

This invention relates to an improved recombinant polyadenylated signal sequence and its use.

The invention provides polynucleotide constructs for the regulation of gene expression by aptamer-based modulation of U1 small nuclear ribonucleoprotein (snRNP)-mediated suppression of polyadenylation and methods of using the constructs to regulate gene expression in response to the presence or absence of a ligand that binds the aptamer. The polynucleotide construct contains a U1 binding site in the context of a riboswitch comprising an effector region and an aptamer such that when the aptamer binds a ligand, target gene expression occurs.

A vector for use in treating cognitive decline and / or memory deficits in a subject in need thereof includes an expression cassette that includes a nucleic acid encoding asprosin and / or an analogue thereof, operably linked to one or more regulatory elements that promote expression of the asprosin and / or an analogue thereof coding sequence, wherein the one or more regulatory elements include a nucleic acid promoter, a polyadenylation (poly(A)) tail signal, and / or posttranscriptional regulatory element.

The present invention describes that a pre-mRNA molecule, which temporarily exists before polyadenylation-site induced cleavage occurs, can be targeted by RNase H recruiting antisense oligonucleotides downstream of a transcripts 3' terminus to reduce expression levels of target genes from which the RNA polymerization originates.

The invention discloses a gene expression enhancing element capable of improving a gene transcription level and an application of the gene expression enhancing element. It is found that a delta satellite region from sweet potato mosaic virus (SPLCV) SBG51 comprises a new DNA sequence capable of remarkably enhancing expression of a reporter gene, the gene expression enhancing effect of the delta satellite region does not change polyadenylation of mRNA, the delta satellite region is independent of a promoter and a terminator, and the space-time mode of the promoter is kept. The invention further finds that AT-rich sequences (such as ATAAA or TTAAA or tandem repeat sequences containing a plurality of ATAAA or TTAAA) are key elements of the above effects, even synthetic AT-rich sequences (AT-rich-1 and AT-rich-2) of 29bp are sufficient to increase the expression of the reporter gene GFP by about 3 times, and the gene expression enhancement effect of the AT-rich sequences is well expressed in both plants and yeast cells. Therefore, the invention can be applied to the aspects of improving the activity of the target gene according to the required level, providing high-level and long-lasting protein production and the like.

A transposon is provided comprising a nucleic acid sequence traversing the chromatin open element (UCOE) as well as a nucleic acid sequence of a gene of interest or a fragment thereof under the control of a promoter and optionally a 3'polyadenylation sequence, both sides of which are Piggybactransposon-specific inverted terminal repeats PB 5 'ITR and PB 3' ITR.

The invention discloses a plasmid vector for stable expression of mammalian cells as well as a construction method and application of the plasmid vector, relates to the technical field of genetic engineering, and discloses a plasmid vector for stable expression of mammalian cells. The plasmid vector comprises a promoter, a polyadenylation signal, an internal ribosome entry site IRES, a replication start site, a selection marker gene and a specific gene; according to the plasmid vector for stable expression of the mammalian cells as well as the construction method and the application of the plasmid vector, an EBNA1 / oriP system is constructed, EBNA1 protein can be specifically combined with an oriP sequence on the plasmid, and the plasmid is firmly adhered to a host cell genome like an anchor and is prevented from being discharged or lost by the cells, so that a target gene can be stably inherited and expressed for a long time along with cell division, and the expression efficiency is improved. The problem of plasmid loss is solved from the source; and the plasmid vector can drive a target gene in various mammalian cells to realize efficient and stable expression of a cross-cell line, and is extremely high in universality.

Provided herein is an expression cassette, the expression cassette comprising: 1) a chimeric promoter, the chimeric promoter comprising an hTERT promoter and a minimum promoter; 2) a coding sequence (CDS); and 3) a poly (A) sequence or a polyadenylation signal sequence. The CDS sequence in the expression cassette is only expressed in a tumor or a cancer cell, or the expression level in the tumor or the cancer cell is far higher than the expression level in a normal cell, so that the expression cassette can be used for treating cancer.

A partially or fully differentiated iPSC-derived microglia cell comprising a transposon comprising a Ubiquitous Chromatin Opening Element (UCOE) nucleic acid sequence, and a nucleic acid sequence of a gene of interest or a fragment thereof and optionally a 3' polyadenylation sequence under control of a promoter, flanked by transposon-specific inverted terminal repeat sequences 5' ITR and 3' ITR.

The invention discloses a single cell 3'end transcriptome variable polyadenylation (APA) analysis method, and belongs to the field of bioinformatics. The analysis process provided by the invention is based on 3'end transcriptome sequencing data of a single cell, combines multiple processes of UMI deduplication, false comparison removal, site merging, internal amplification removal, polyA site removal with few supporting cells, site verification and the like, and can realize efficient detection and quantification of APA at the single cell level. The method is especially suitable for single cell APA analysis of single cell 3'end transcriptome sequencing data, including 3 'end single cell sequencing data after 10x scRNA-Seq cell subset combination, 3' end single cell sequencing data based on a C1 chip, and even full-length transcriptome data.

The present invention relates to a recombinant oleaginous yeast cell having at least one CEX1 gene disruption, comprising chimeric gene constructs, each of said chimeric gene constructs comprising a yeast-expressible promoter and a 3' end region comprising transcription termination and polyadenylation signals functioning in a yeast cell, wherein said chimeric gene constructs allow for overexpression of DGA1, ACC1, MCE2, and ACL genes, said ACL gene being ACL1 and / or ACL2 gene. In a second aspect, the invention relates to a method for producing lipids using said transformed oleaginous yeast cell. The invention further pertains to lipids obtainable by the method, a composition comprising said lipids, a composition comprising transformed oleaginous yeast cell, and a food product obtainable therefrom.

Disclosed herein are gene therapy vectors for efficient transduction of cells to express human beta globin genes. Specifically disclosed is an expression vector comprising: an expression cassette of a beta globin gene comprising an exon and an intron of a human beta globin gene, and a cis-acting element comprising one or more of WPRE, an SV40 polyadenylation signal and / or SV40 ori. The disclosed expression vectors have significantly enhanced viral vector packaging efficiency in viral vector packaging cell lines, which results in efficient integration of lentiviral vectors in target cells and high expression levels of beta globin genes. Pharmaceutical compositions and methods of treatment utilizing such expression vectors are also disclosed.

The invention provides polynucleotide constructs for the regulation of gene expression by aptamer-based modulation of U1 snRNP-mediated suppression of polyadenylation and methods of using the constructs to regulate gene expression in response to the presence or absence of a ligand that binds the aptamer. The polynucleotide construct contains a U1 binding site in the context of a riboswitch comprising an effector region and an aptamer such that when the aptamer binds a ligand, target gene expression occurs.

The present invention relates to a nucleic acid sequence, comprising or coding for a coding region, encoding at least one peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, at least one histone stem-loop and a poly(A) sequence or a polyadenylation signal. Furthermore the present invention provides the use of the nucleic acid for increasing the expression of said encoded peptide or protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine, e.g. for use in the treatment of infectious diseases. The present invention further describes a method for increasing the expression of a peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal.

In one aspect, the invention provides a method of screening a human subject to determine if said subject has a genetic predisposition to develop, or is suffering from Facioscapulohumeral Dystrophy (FSHD), said method comprising: (a) providing a biological sample comprising genomic DNA from the subject; and (b) analyzing the portion of the genomic DNA in the sample corresponding to the distal D4Z4-pLAM region on chromosome 4 and determining the presence or absence of a polymorphism resulting in a functional polyadenylation sequence operationally linked to exon 3 of the DUX4 gene.

The present invention relates to improved recombinant polyadenylation signal sequences and use thereof.

The application discloses a variable polyadenylation dynamic analysis method and device, and relates to the technical field of gene analysis. The method comprises the following steps: acquiring initial spatial transcriptome data and initial spatial bright field images; performing first preprocessing on the initial spatial transcriptome data to obtain target spatial transcriptome data; performing second preprocessing on the initial spatial bright field images to obtain target spatial bright field images; performing data analysis and integration processing to obtain spatial variable polyadenylation data, spatial gene expression data and spatial cell distribution dynamic data; constructing a gene expression regulation network; generating integration analysis results; performing space-time analysis to obtain space-time analysis results; performing trajectory analysis to obtain trajectory analysis results; performing disease progression analysis to obtain disease progression analysis results; and generating variable polyadenylation dynamic analysis results. The application realizes variable polyadenylation dynamic analysis, improves accuracy, and provides multi-angle biological understanding. The application can be applied to the technical field of gene analysis.

The invention provides a method of regulating U1 activity associated with its splicing role as well as its role in protecting pre-mRNAs from premature termination by cleavage and polyadenylation, thereby modulating expression of a gene or genes. In one embodiment, the invention includes compositions and methods for regulating gene expression and treating diseases associated with dysregulated gene expression.

In one aspect, the invention provides a method of screening a human subject to determine if said subject has a genetic predisposition to develop, or is suffering from Facioscapulohumeral Dystrophy (FSHD), said method comprising: (a) providing a biological sample comprising genomic DNA from the subject; and (b) analyzing the portion of the genomic DNA in the sample corresponding to the distal D4Z4-pLAM region on chromosome 4 and determining the presence or absence of a polymorphism resulting in a functional polyadenylation sequence operationally linked to exon 3 of the DUX4 gene.

One-way SV40 late polyadenylation signals and combinations of such one-way SV40 late polyadenylation signals with other polyadenylation signals, such as bovine growth hormone (BGH) polyadenylation signals, are provided. The polyadenylation signal can be used in nucleic acid constructs and compositions that allow for insertion of a coding sequence for a polypeptide of interest into a target genomic locus and / or expression of a polypeptide of interest. The nucleic acid constructs and compositions are useful in methods for integrating a coding sequence of a polypeptide of interest into a target genomic locus and methods for expressing a polypeptide of interest in a cell.

This invention proposes a spatial whole-transcriptional sequencing method and application suitable for tobacco petiole tissue. The method employs tissue embedding and frozen sectioning techniques suitable for tobacco petioles, effectively ensuring uniform and flat sections and maintaining structural integrity. A pretreatment solution suitable for plant tissue permeation effectively enzymatically dissolves plant cell walls and increases cell membrane permeability, facilitating better RNA release. An in-situ polyadenylation treatment method suitable for tobacco petiole tissue effectively polyadenylates non-coding RNA in the sectioned samples, improving non-coding RNA capture efficiency. A method suitable for spatial whole-transcriptional sequencing workflows to remove rRNA from cDNA libraries effectively removes rRNA from the library through wet experimental ends, avoiding reduced sequencing depth for other RNA types due to excessive ribosomal RNA capture, thus greatly improving data utilization. This invention has broad application prospects.