35 results about "Alternative splicing" patented technology

The invention discloses a composition, a detection method and a kit for SpaceAS variable splicing space in-situ detection of nucleic acid. The composition comprises a pi-type probe, a U-type probe, an O-type probe, an S-type probe and a signal probe, and the O-type probe adopts an in-vitro cyclization strategy, so that the problem of low hybridization efficiency caused by dependence on enzymatic cyclization reaction in a traditional hybridization method is effectively solved; meanwhile, an efficient probe hybridization system, a high-sensitivity signal amplification mechanism and a multi-round iterative imaging process are combined, so that the detection sensitivity and specificity are remarkably improved. The detection method has the advantages of high efficiency, high sensitivity, high specificity, good repeatability, strong expandability and the like through the synergistic effect of five-step single-stranded DNA probe hybridization reaction and one-step DNA amplification reaction, and can be widely applied to space in-situ detection and analysis of variable splicing events.

A method and compound for regulating SCN1A gene mRNA and protein expression. Provided is a use of a guide agRNA in preparation of a drug for altering expression of a target protein in a cell of a subject. The cell has a precursor mRNA encoding the target protein, wherein the precursor mRNA comprises an intron, an exon flanking a 5' splice site of the intron, and an exon flanking a 3' splice site or a pseudo 3' splice site of the intron; the guide agRNA is capable of binding to the precursor mRNA to form a structure capable of recruiting ADAR in the cell, thereby forming a complex containing an ADAR protein; and the target protein is a Nav1.1 protein. By regulating alternative splicing of a target gene pre-mRNA, the functional RNA of the target gene and the function or expression of the target protein are affected, thereby achieving the objective of disease treatment.

The present invention relates to methods and devices for identification and quantification, including low abundance nucleotide base mutations, insertions, deletions, translocations, splice variants, miRNA variants, alternative transcripts, alternative start sites, alternative coding sequences, alternative non-coding sequences, alternative splicing, exon insertions, exon deletions, intron insertions, or other rearrangements and / or methylated nucleotide bases at the genomic level.

The application provides an antisense oligonucleotide Jag-i9 ASO and application thereof, relates to the technical field of biological medicine, and the antisense oligonucleotide Jag-i9 ASO has the sequence of 5'-ACTGGGCCCTGCACCTGA-3'. By providing the antisense oligonucleotide of a specific sequence, the binding site of target heterogeneous ribonucleoprotein K and the Jag2 gene is targeted, the expression of the pro-inflammatory Jag2 subtype is inhibited, the Jag2 gene splicing site can be accurately targeted, the heterogeneous ribonucleoprotein K binding function is specifically blocked, a targeted intervention means is provided for the treatment of myocardial ischemia-reperfusion injury, the generation of the pro-inflammatory subtype is inhibited by efficiently and accurately regulating the alternative splicing of a specific gene, myocardial cell apoptosis and inflammatory response are reduced, myocardial function is improved, and a new approach is provided for the prevention and treatment of myocardial ischemia-reperfusion injury, the treatment effect is improved, and the advantages of reducing side effects are achieved.

The present invention provides a mesenchymal stem cell having blood compatibility, and a preparation method and use thereof, the mesenchymal stem cell inducing exon skipping of an exon 5 by performing gene modification on the exon 5 of an F3 gene, thereby increasing the expression of a selective splicing tissue factor (asTF), and improving the blood compatibility of the mesenchymal stem cell. The present invention relates to a method for inhibiting thrombosis by reducing or inhibiting the expression or activity level of a full-length tissue factor (flTF), which is a blood coagulation promoter, and thereby inhibiting thrombosis.

The Quadruplet Expanded DNA (QED) eukaryote genetic code comprising twenty nondegenerate QED codons encode proteins (the protein-encoding codons), and thirty-five nondegenerate QED codons (the noncoding codons) being highly correlated with cis-regulatory elements control and regulate transcription, alternate splicing, and polymerization in eukaryotic protein synthesis using canonical amino acids. The QED eukaryote genetic code is an advancement to gene therapeutics that allows for the correction of dysfunctional proteins. Additionally, the QED eukaryote genetic code is further applicable for changing paradigms relating to identifying cures for monogenic rare, multigene cancer, and neurodegenerative diseases.

A method for regulating the mRNA and protein expression of an SCN1A gene, and a compound. Provided is a use of an antisense oligonucleotide guide RNA in the preparation of a drug for altering the expression of a target protein in a cell of a subject. The cell has a pre-mRNA encoding the target protein, wherein the pre-mRNA comprises an intron, an exon flanking a 5' splice site of the intron, and an exon flanking a 3' splice site or pseudo 3' splice site of the intron; the antisense oligonucleotide guide RNA can bind to the pre-mRNA to form a structure capable of recruiting ADAR in a cell, thereby forming a complex containing an ADAR protein; and the target protein is an Nav1.1 protein. That is, the function or expression of the functional RNA and target protein of a target gene is affected by regulating the alternative splicing of the pre-mRNA of the target gene, thereby achieving the objective of treating diseases.

A variable splicing event in a gene can result in a non-productive mRNA transcript, which in turn can result in abnormal protein expression, and a therapeutic agent that can target a variable splicing event in a gene can modulate the expression level of a functional protein and / or inhibit abnormal protein expression in a patient. Such therapeutic agents are useful for the treatment of conditions or diseases caused by protein deficiency and / or mitochondrial function deficiency.

The application provides application of a selective splicing factor SF3A2 as a biomarker in preparation of a reagent for breast cancer diagnosis, treatment, prognosis evaluation and chemotherapy sensitivity evaluation. The application provides application of a detection reagent for expression level of a selective splicing factor SF3A2 gene or protein in preparation of a kit for breast cancer diagnosis, prognosis evaluation and chemotherapy sensitivity evaluation. Further, the application provides application of an expression inhibitor for a selective splicing factor SF3A2 gene or protein in preparation of a kit for breast cancer treatment and chemotherapy sensitivity improvement.

Provided herein are chimeric transactivator minigenes, where the alternative splicing of the minigene determines whether a transactivator is expressed. Expression of the transactivator results in the transcription of a target gene that is under the control of a designer promoter sequence. Alternatively, provided herein are chimeric target gene minigenes, wherein the alternative splicing of the minigene directly determines whether the target gene is expressed. The target gene may encode an inhibitory RNA, a CRISPR-Cas9 protein, or a therapeutic protein.

The application belongs to the field of genetic engineering, and particularly relates to a polynucleotide construct capable of expressing LMNA and application thereof in preventing and treating DCM. The synthetic intron in the application comprises: I) a nucleotide sequence as shown in SEQ ID No. 8, II) a nucleotide sequence having at least one mutation in the 1-8th position of the sequence shown in SEQ ID No. 8, or a nucleotide sequence having at least 90% homology with the sequence shown in I) or II). By using the synthetic intron in the application, two proteins can be simultaneously expressed through alternative splicing, and the ratio of the two proteins or the expression of a single protein can be regulated through mutation of the splicing signal. Compared with the existing gene therapy method for dilated cardiomyopathy caused by LMNA mutation, the expression vector for simultaneously expressing Lamin A and Lamin C proteins in the application has stronger rescue effect.

An infection status of a subject is determined using sequence reads from a biological sample of the subject. For each respective alternative splicing (AS) event in a plurality of AS events, there is determined (i) a corresponding first abundance metric of the AS event in the biological sample based on a mapping of each sequence read to one or more reference splice junctions in a plurality of reference splice junctions, and (ii) a corresponding second abundance metric of the AS event in the biological sample based on a mapping of each respective sequence read of the plurality of sequence reads to one or more reference isoforms in a plurality of reference isoforms. Each AS event corresponds to a locus in a reference genome. The first and second abundance metrics for each AS event are inputted into a model to obtain a predicted infection status of the subject as model output.

The present application relates to a method for finding new variable splicing events in three-generation full-length transcript sequencing data in the field of bioinformatics. The method for detecting new variable splicing events in three-generation full-length transcript sequencing data of a test sample of the present application comprises the following steps: 1) obtaining three-generation full-length transcript sequencing data of a test sample; 2) performing gene function annotation on the three-generation full-length transcript sequencing data of step 1) to obtain transcript set 1, wherein the transcript set 1 comprises NNC and NIC; 3) screening whether variable splicing events occur in the transcript set 1 of step 2) to obtain transcript set 2 with variable splicing events; 4) filtering and screening the transcript set 2; 5) after step 4) is completed, screening new splicing isoforms formed by known variable splicing events and new splicing isoforms formed by multiple variable splicing events; 6) after step 5) is completed, outputting a variable splicing event result information file.

The invention provides antisense oligonucleotide for regulating and controlling IL1RL2 gene variable splicing as well as a preparation method and application of the antisense oligonucleotide. Specifically, the invention provides an antisense oligonucleotide which interferes with variable splicing of an IL1RL2 gene, thereby inducing the generation of a truncated or concealed exon insertion type IL1RL2 splicing isomer transcript. The antisense oligonucleotide therapy provided by the invention can reduce the proportion of mRNA for coding full-length functional IL-36R and realize activation and blocking of an IL-36R signal channel by inducing splicing jump of an exon 4 or an exon 10 of an IL1RL2 gene precursor mRNA or splicing insertion of a hidden exon sequence of an intron 8. The compound can be used for treating autoimmune diseases, allergic diseases, neurodegenerative diseases or cancers related to abnormal activation of IL-36R.

Alternative splicing events in genes can lead to non-productive mRNA transcripts which in turn can affect protein expression level, and therapeutic agents which can target the alternative splicing events in genes can modulate the expression level of functional proteins in patients and / or inhibit aberrant protein expression. Such therapeutic agents can be used to treat a condition or disease caused by protein deficiency.

Provided herein are methods for treating conditions and diseases characterized by SCN1A, SCN8A, or SCN5A protein deficiency by targeting variable splicing events in the SCN1A gene and modulating expression levels of functional proteins and / or inhibiting aberrant protein expression in Delavir Syndrome patients.

Provided is an antisense oligonucleotide which interferes with the alternative splicing of an IL1RL2 gene, thereby inducing the production of a truncated IL1RL2 splice isoform transcript. The antisense oligonucleotide can reduce the proportion of mRNA encoding full-length functional IL-36R by means of inducing exon 4 skipping in the pre-mRNA of the IL1RL2 gene, thereby blocking the activation of an IL-36R signaling pathway, and can be used for treating autoimmune diseases, allergic diseases, neurodegenerative diseases or cancers associated with abnormal activation of IL-36R.

The present invention relates to efficient splice-conversion antisense oligonucleotides that modulate selective splice of a precursor mRNA encoding an advanced glycosylation end product receptor (RAGE) or a portion thereof, and methods of using the antisense oligonucleotides to treat RAGE-related disorders. In one aspect, the present invention provides an antisense oligonucleotide (AON) consisting of the nucleotide sequence TGCCGCCTTGCCACAAG (5 '-3', SEQ ID NO: 1) or GCCGCCTTGCCACAAGA (5 '-3', SEQ ID NO: 2), preferably wherein the antisense oligonucleotide comprises at least one modified nucleotide.

The application discloses a screening method for an early diagnosis marker of Alzheimer disease based on alternative splicing, and relates to the technical field of biomedical diagnosis. The application aims to solve the problems of single feature dimension, insensitivity to splicing changes, insufficient robustness and the like in the prior screening method for an early diagnosis marker of Alzheimer disease, and provides a multi-dimensional feature fusion screening method based on alternative splicing.

This invention relates to the field of biomedicine, specifically to a synergistic and targeted epigenetic pharmaceutical composition for the treatment of lung cancer. In studies of drug-resistant lung cancer cells, it was found that overexpression of the NSUN7 gene in drug-resistant lung cancer cells can inhibit MZF1. L The expression of MZF1 also promotes S When the expression of NSUN7 gene was suppressed, no promotion of MZF1 was found. L The expression of MZF1 can be suppressed at the same time. S The expression of NSUN7 gene was inhibited, meaning no opposite result was obtained. However, when NSUN7 gene expression was inhibited and EGFR-TKI inhibitors were administered to drug-resistant lung cancer cells, MZF1 cells in drug-resistant lung cancer cells showed a positive result. L Upregulation, MZF1 S Decreased expression of NSUN7 leads to the transformation of drug-resistant lung cancer cells into drug-sensitive cells. Therefore, interfering with NSUN7 in combination with epigenetic drugs to regulate the alternative splicing of methylation-sensitive transcription factors can increase the specificity of regulatory targets and synergistically enhance the in vitro and in vivo antitumor effects.

PI3Kδ implicates hematologic cancers and solid tumors. Alternative splicing is a post-transcriptional process for acquiring proteomic diversity in eukaryotic cells. Emerging evidence highlights the involvement of aberrant mRNA splicing in cancer development / progression. PI3Kδ-L and PI3Kδ-S are overexpressed in advanced solid tumors, such as prostate, breast, colon, lung and pancreatic cancers. Differential PI3Kδ and PI3Kδ-S expression profiles were identified in a panel of solid tumor cells. PI3Kδ inhibitor Idelalisib and SRPK1 / 2 inhibitor SRPIN340 were employed to assess their efficacies on inhibiting the PI3Kδ-expressing solid tumors. Idelalisib effectively inhibits PI3Kδ-L and its downstream signaling. Idelalisib fails to inhibit PI3Kδ-S activity and its downstream signaling. SRPIN340 reverses the aberrant mRNA splicing, thereby inhibiting the downstream AKT / mTOR signaling. In vitro functional assays further demonstrate that a combination of Idelalisib and SRPIN340 achieve a synergistic drug effect, with drastically reduced cell viabilities / growths of tumor spheroids, in inhibiting the advanced tumor cells.

The application relates to the technical field of gene analysis, and provides an Alzheimer disease gene prediction method and equipment based on alternative splicing, which comprises the following steps: performing difference analysis on the expression amount of all genes and the expression amount of transcripts to obtain expression amount difference characteristics, expression amount difference characteristics and expression proportion difference characteristics of the genes, and performing difference analysis on alternative splicing events of all sample gene signals to obtain alternative splicing difference characteristics of each gene; splicing the expression amount difference characteristics, the transcript expression amount difference characteristics, the expression proportion difference characteristics and the alternative splicing difference characteristics of each gene to obtain final characteristics of each gene; constructing a function correlation network according to the final characteristics of all genes; and performing gene disease prediction on the function correlation network to obtain the probability that each gene is related to Alzheimer disease. The method can improve the accuracy and reliability of Alzheimer disease gene prediction.

The present invention relates to potent antisense oligonucleotides that modify splicing, modulating the alternative splicing of pre-mRNA encoding the Receptor of Advanced Glycation End Products (RAGE) or a portion thereof, and methods for treating RAGE-related disorders using such antisense oligonucleotides. In one aspect, the present invention provides an antisense oligonucleotide (AON) consisting of the nucleotide sequence TGCCGCCTTTGCCACAAG (5'-3'; SEC ID NO: 1); or GCCGCCTTTGCCACAAGA (5'-3'; SEC ID NO: 2), preferably wherein the antisense oligonucleotide comprises at least one modified nucleotide.

This disclosure provides a DNA construct comprising: (a) a start exon encoding a start codon; (b) a first frameshift exon downstream of the start exon, the first frameshift exon including a first receptor splice site, wherein inclusion or exclusion of the first frameshift exon in alternative splicing determines the downstream reading frame of a mature mRNA transcript; and (c) a first specific exon downstream of the first frameshift exon, the first specific exon including a second receptor splice site, a first reporter gene, and a second reporter gene, wherein the first reporter gene is located upstream or downstream of the second reporter gene, and wherein the first reporter gene and the second reporter gene are in different reading frames, wherein exclusion of the first frameshift exon causes the first reporter gene to be in the same reading frame as the start codon, and wherein inclusion of the first frameshift exon causes the second reporter gene to be in the same reading frame as the start codon.

The invention relates to the technical field of gene engineering, in particular to application of a CmESI gene in regulating and controlling dwarfing of a muskmelon plant. According to the invention, methods of genetic analysis, mixed pool sequencing, Mutmap and Kasp genotype analysis and the like are combined to locate a gene causing extreme dwarfing of a plant, an SNP variation site of G / A existing in a first basic group of a seventh exon of a CmESI gene in a mutant esi is found out, the variation site causes CmESI to generate a variable shearing form with a retained sixth intron, and the mutation site is used for detecting the extreme dwarfing of the plant. A termination codon is formed in advance and cannot be translated into complete protein, so that the CmESI gene loses functions; the expression of CmESI in the 13C germplasm is inhibited by using a virus-induced gene silencing technology, and the plant height of a TRSV-CmESI plant is obviously lower than that of a control group plant; therefore, the muskmelon CmESI gene has certain theoretical significance and practical value in regulating and controlling the height of muskmelon plants.

The present invention relates to a method of modulating selective splicing of a precursor mRNA encoding an advanced glycosylation end product receptor (RAGE) or a portion thereof using an antisense oligonucleotide (AON) to modify the expression and / or activity of RAGE isoforms. In one aspect, the invention provides a method for promoting the production of endogenous soluble RAGE and / or reducing the production of membrane-bound RAGE (mRAGE) in a subject, the method comprising: in a first administration, administering to the respiratory tract of the subject an antisense oligonucleotide (AON) that promotes the production of endogenous soluble RAGE and / or reduces the production of membrane-bound RAGE (mRAGE), in a second administration, AON that promotes the production of endogenous soluble RAGE and / or reduces the production of membrane-bound RAGE (mRAGE) is administered to the respiratory tract of the subject, where the second administration is at least about 7 days after the first administration, thereby promoting the production of endogenous soluble RAGE and / or reducing the production of membrane-bound RAGE (mRAGE) in the subject.

The application belongs to the technical field of biology, and discloses a 5'UTR sequence variable splicing analysis method, device, equipment and medium; the method comprises the following steps: locating the coordinates of 5'UTR sequences in conventional transcriptome data; comparing the 5'UTR sequences with reference genome data, and performing multiple screening on the 5'UTR sequences that occur variable splicing in the conventional transcriptome data according to the comparison result to obtain screened variable splicing 5'UTR sequences; and performing RNA secondary structure analysis on the screened variable splicing 5'UTR sequences to obtain the RNA secondary structure of the screened variable splicing 5'UTR sequences; the application realizes accurate 5'UTR variable splicing analysis by using conventional transcriptome data and reference genome, and can be suitable for model species and non-model species, and provides a useful tool for studying gene expression regulation, especially the relationship between 5'UTR variable splicing and gene expression regulation.

The application discloses a SNP molecular marker for affecting the number of 210-day egg production of chickens by affecting variable splicing of a CAPN6 gene and application thereof, belongs to the field of molecular genetics and agricultural animal breeding, and discloses that the SNP molecular marker is located at the position of 13053733 bp of chicken chromosome 4, the reference genome version is GRCg7b, has T / C polymorphism, and the individual with the genotype CC at the position has a higher number of 210-day egg production than the individual with other genotypes. The application constructs a molecular marker system which is significantly related to splicing expression of CAPN6 and can be used for early prediction and selection of the number of 210-day egg production of chickens, and provides a precise and efficient molecular technical path for breeding of laying hens. The application has important significance for promoting molecular marker assisted breeding of reproductive traits of laying hens.

Described herein are nucleic acids and recombinant viral vectors comprising said nucleic acids, wherein said nucleic acids comprise an interferon that is regulated by alternative splicing.

This disclosure relates to nucleic acids for use in regulating the transcription of linked transgenes by an exon inclusion mechanism. In certain embodiments of this disclosure, a nucleic acid molecule is provided comprising a minigene located adjacent to the 5' side of a transgene, wherein the minigene comprises (a) a first exon adjacent to the 5' side of a first intron, (b) a start codon, and (c) a splice regulator binding site, the splice regulator binding site comprising the nucleic acid sequence DGAGTDDGHV (SEQ ID NO: 82) or DGAGTDDNHV (SEQ ID NO: 83), where D is A, G, or T, R is A, G, N is A, C, G, or T, H is A, C, or T, and V is A, G, or C.