97 results about "Alternative splicing" patented technology

To provide a medicine for treating a disease in a mammal.SOLUTION: Provided is a medicine for treating a disease in a mammal, comprising an expression cassette that is a first expression cassette comprising a chimeric gene operably linked to a first expression control element, the chimeric gene comprising a first portion comprising a minigene having an alternatively spliced exon and a second portion encoding a first RNA encoding a protein, the first portion being 5' to the second portion, the second portion lacking an initiation codon or start codon, the alternatively spliced exon comprising a translation initiation regulatory sequence, expression of the protein being controlled by inclusion of the alternatively spliced exon, and the medicine being used in combination with a small molecule splicing modifier that modulates splicing of the alternatively spliced exon.SELECTED DRAWING: Figure 1

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

Agents that target a processed mRNA, e.g., the 5′ UTR of the processed mRNA, can modulate protein expression, e.g., via modulation of translation of the processed mRNA. Alternative splicing events in genes can lead to non-productive mRNA transcripts which in turn can lead to aberrant protein expression. Agents that target the alternative splicing events in genes can modulate the expression level of proteins. Therapeutic agents, which can modulate protein expression by targeting a processed mRNA and / or alternative splicing events, can promote functional protein expression in patients and / or inhibit aberrant protein expression. Such therapeutic agents can be used to treat a condition or disease associated with protein deficiency and / or mitochondrial function deficit.

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.

The present invention provides an agent capable of modulating variable splicing in the intron of the SLC20A2 gene, and its use for modulating the expression of functional PiT2 protein to prevent and / or treat disease. The invention further provides a brain calcification model based on variable splicing in the intron of the human SLC20A2 gene, a preparation method of the brain calcification model, and application of the model to indication and analysis of the incidence stage of brain calcification, or to a model for researching phosphorus homeostasis imbalance in brain or a target of brain calcification, or to screening of drugs.

The present invention provides a versatile platform system for regulating gene expression in cells of eukaryotic organisms, including plants, algae and fungi, and mammals. A platform system of the invention includes an expression control element and expression system comprising the same. Expression control elements according to the invention can be inserted into a variety of cell types having the capability of alternative splicing and can be regulated by a polyamine or polyamine analogues. Host cells comprising the expression control elements of the invention are typically grown in a cell or tissue culture, or form part of a living plant, algae, or fungi.

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.

The present invention belongs to the field of biochemistry and discloses a polypeptide comprising: (I) an amino acid sequence as shown in SEQ ID No. 1; or (II) an amino acid sequence formed by adding one or more amino acids before or after the amino acid sequence as shown in (I); or (III) a cell-penetrating polypeptide obtained by combining the polypeptide shown in (I) or (II) with a cell-penetrating peptide. The polypeptide significantly inhibits the growth and proliferation of two prostate cancer cell lines (DU145 and PC3), as well as RNA alternative splicing. The present invention also discloses a drug containing the polypeptide and its uses.

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

The present invention relates to a series of antisense oligonucleotides containing morpholine 3- thiophosphoramidate nucleotides or morpholine 3-thiophosphoramidate and DNA / RNA (modified) nucleotides having phosphorothioates (pS) linkers. They are specifically designed to be complementary to nucleic acid sequences within the Signal Transducer and Activator of Transcription 3 (STAT3) pre-mRNA or mature sequences, leading to the degradation of STAT3 mRNA or inhibition of STAT3 translation, or induce alternative splicing of STAT3 resulting in an increase of STAT3β isoform relative to STAT3α isoform. The antisense oligonucleotides are used for the treatment of diseases that could be modulated by the STAT3 pathway, which has been implicated in tumor initiation, progression, metastasis, drug resistance and immune suppression. As a result, these novel antisense oligonucleotides could be used for the treatment of STAT3 related diseases, such as cancer or autoimmune diseases.

The present invention discloses the use of sIL-36R, DDX5, and DDX5 / sIL-36R in the preparation of drugs for inhibiting, alleviating, reducing, and / or treating atopic dermatitis and / or psoriasis. The present invention also discloses the biomarker DDX5 and / or sIL-36R and / or DDX5 / sIL-36R for psoriasis and / or atopic dermatitis and their applications. DDX5 is downregulated in keratinocytes in the skin lesions of psoriasis and / or atopic dermatitis, but the expression of DDX5 returns to normal levels after the skin lesions subside. The low expression of DDX5 leads to changes in the alternative splicing of IL-36R pre-mRNA in keratinocytes, resulting in an increase in IL-36R and a decrease in soluble sIL-36R, thereby promoting the disorder of the IL-36R signaling pathway and inducing skin inflammation; while injecting sIL-36R can effectively inhibit the IL-36R signaling pathway in atopic dermatitis mice or psoriasis mice, and inhibit the expression of cytokines in the skin lesions of atopic dermatitis or psoriasis mice to alleviate atopic dermatitis. The sIL-36R, DDX5, and DDX5 / sIL-36R can be used as targets and biomarkers for the diagnosis and treatment of inflammatory skin 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 invention discloses a pituitary tumor subtype distinguishing method which comprises the following steps: acquiring a pituitary tumor subtype sample, extracting RNA (Ribonucleic Acid) of a sample tissue and reversely transcribing the RNA into cDNA (Complementary Deoxyribonucleic Acid); through PCR amplification of a variable splicing gene primer and in combination with an agarose gel electrophoresis experiment, a standard screen band of a gene having a specific splicing event in a corresponding pituitary tumor subtype is obtained; carrying out PCR (Polymerase Chain Reaction) amplification and agarose gel electrophoresis experiments on a pituitary tumor sample to be detected by utilizing the gene to obtain an experimental screen strip of the pituitary tumor sample to be detected; and comparing the experimental screen strip with a standard screen strip presented by the corresponding pituitary tumor subtype, and distinguishing the subtype of the pituitary tumor sample to be detected. According to the invention, effective subtype diagnosis can be carried out in an operation, a subtype diagnosis effect can be achieved by utilizing a smaller tissue, diagnosis identification can be carried out by quickly utilizing a postoperative tissue sample, and if part of antibodies are out of order or poor in sensitivity, pathological diagnosis can be supplemented to diagnose part of pituitary tumor subtypes which cannot be determined temporarily.

A targeted high-throughput sequencing method for detecting splice isoforms, including a method for establishing a sequencing library for high-throughput sequencing, comprising the following steps: 1) reverse transcription of sample RNA using a reverse transcription primer, adding common dNTPs and 3'-modified dNTPs to generate a first-strand cDNA; 2) ligating an oligonucleotide fragment with an alkyne modification at the 5' end to the cDNA fragment obtained in step 1) via a click chemistry reaction; 3) performing PCR amplification using the reaction product of step 2) as a template; and 4) obtaining a sequencing library. Targeted enrichment primers can be introduced during the reverse transcription or PCR amplification steps. The enrichment step reaction system contains multiple gene-specific primers, each designed based on the downstream exon segment of the alternative splicing event in the transcript, ensuring that the random-length fragments generated by reverse transcription cover the splice site.

Alternative splicing events in genes can lead to non-productive mRNA transcripts which in turn can lead to aberrant or reduced protein expression, and therapeutic agents which can target the alternative splicing events in the 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.

The present invention discloses a leucine-rich repeat protein kinase family gene GhLRRK1 and its application. The genomic sequence of the leucine-rich repeat protein kinase family gene GhLRRK1 is SEQ ID NO.2. In the upland cotton population, by combining whole-genome resequencing and transcriptome data of ovules one day after flowering, genome-wide association analysis and alternative splicing locus analysis were carried out, and a gene GhLRRK1 significantly associated with cotton lint percentage was identified. There is an alternative splicing site regulated by SNP on this gene. The SNP site has two haplotypes, A and G. Intron retention occurs at the alternative splicing site in the G haplotype, and the intron is not retained in the A haplotype. The lint percentage of the G haplotype is significantly higher than that of the A haplotype, and it may be a causal gene regulating the cotton lint percentage trait. The present invention provides important research value and application prospects of this gene in the efficient identification of upland cotton varieties with high lint percentage, the improvement and cultivation of new cotton varieties with high lint percentage.

The invention discloses an application of a YTHDC1 gene in regulating myoblast proliferation or differentiation and skeletal muscle development. The influence of the YTHDC1 gene on proliferation and differentiation of myoblasts is researched by inhibiting and overexpressing the YTHDC1 gene in the myoblasts, and it is found that the overexpressed YTHDC1 gene has the functions of promoting differentiation of the myoblasts and inhibiting proliferation of the myoblasts; myoblast proliferation can be promoted and differentiation of myoblasts can be inhibited by inhibiting YTHDC1 gene expression, and muscle growth of mice after birth can be promoted. And further research finds that the YTHDC1 gene affects muscle development by regulating and controlling the variable splicing process of muscle development related genes such as AkaP13, Smarca2, Tnnt3 and Neb. Experimental results provide a research basis for skeletal muscle growth and development regulation and control, and also provide theoretical support for improving meat product quality.

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.

Aspects of the disclosure relate to the discovery that antisense oligonucleotides (ASOs) that bind to single regions of a CACNA1A pre-mRNA to affect alternative splicing lead to an efficient modulation of protein produced from matured CACNA1A pre-mRNA. This modulation, which can be as an increase or decrease, can alleviate issues related to gain-of-function or loss-of-function mutations in the CACNA1A gene, respectively, including those that lead to haploinsufficiencies. Alleviating such issues can be useful to treat neurological disorders that result from such mutations.

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 invention relates to negative functional modulators of at least one variant of the non-erythrogenic erythropoietin (V-EPO) and pharmaceutical compositions or kits containing them. Such functional negative modulators of V-EPO may be a mono- or multi-specific antibody anti-EV3, anti-EV4, anti-EV1-4, anti-EV1-5, anti-EV1-1, or EV2-1, or anti-Epo receptor (EpoR) anti-EPHB4, anti-CSF2RB, an antisense oligonucleotide, DNA decoy, RNA decoy, a ribozyme, an antagomiR, a shRNA, LNA or siRNA.Several uses of these functional modulators are described, which have been advantageously employed as a medicament and for the treatment of an oncological pathology, a proliferative pathology, chronic inflammatory diseases on an autoimmune and non-autoimmune basis, of neurodegenerative diseases, and in the treatment of patients undergoing organ or tissue transplantation.The invention also describes variants of EPO for use in the diagnosis and in the treatment of an oncological pathology, a proliferative pathology, chronic inflammatory diseases on an autoimmune and non-autoimmune basis, of neurodegenerative diseases, and in the treatment of patients undergoing an organ or tissue transplantation and as a diagnostic agent.According to another aspect, a monoclonal antibody to at least one of the variants of the erythropoietin is described.According to yet another aspect, the use of at least one alternative splicing variant of non-erythrogenic EPO and the measurement thereof at tissue and / or systemic level is described, as well as the study of the methylation status of the promoters of the genes involved in the EPO signalling pathway (by way of example EPO, EPOR, EPHB4, CSF2RB), as diagnostic, prognostic and predictive markers of an oncological pathology, a proliferative pathology, neurodegenerative or inflammatory pathology.

The present invention belongs to the technical field of epigenetic markers associated with bull sperm motility, and specifically relates to an epigenetic marker for screening and / or detecting high and low sperm motility in bulls and its application. The present invention focuses on the differences in DNA methylation in the sperm genomes of full-sib Holstein bulls with high and low sperm motility and their functions. Simplified methylation sequencing and transcriptome sequencing were performed on the genomic DNA and total RNA of the sperm, respectively, to explore the molecular mechanism by which DNA methylation plays a role in bull sperm motility. It was determined that high methylation levels in the exon 29 region of the PBRM1 gene are associated with high sperm motility in bulls. It was further determined that the methylation levels of the exon 29 region of the PBRM1 gene regulate gene expression and function by altering alternative splicing patterns, laying the foundation for screening bulls with high sperm motility and for genetic breeding of dairy cows, and therefore has excellent practical application value.

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.

A therapeutic composition is provided which comprises at least one agent which specifically interferes with PTBP2-binding in the SYNGAP1 gene region to prevent dysfunctional protein production caused by an alternative splicing event, which dysfunctional protein is associated with a disease or disorder. The agent can be an anti-sense oligonucleotide, an RNAi, or combinations thereof. The composition may further comprise a pharmaceutically acceptable aqueous diluent suitable for intrathecal injection. Also provided are methods of treating SYNGAP1-related neurodegenerative disorders.

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.