56 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.

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.

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.

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 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 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.

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 recombinant CAR-NK cells or CAR T-cells, expressing a CAR binding to the antigen CLEC12A or a functional alternatively spliced transcript variant thereof, wherein at least one immune checkpoint receptor protein, such as, for example NKG2A, CLEC12A, PD-1, TIM-3, TIGIT and / or KIRS, is inactivated. These highly functional immune checkpoint-inactivated CAR-NK cells or CAR T-cells target cancer-associated antigens or are adapted for a treatment of autoimmune diseases. Furthermore, the present invention relates to a non-virus-based method for producing a CAR-NK cell or CAR T-cell expressing an antigen-targeting chimeric antigen receptor (CAR) and a recombinant CAR-NK cell or CAR T-cell as produced, in particular a CAR- NK cell or CAR T-cell targeting the cancer-associated antigen CLEC12A. The present invention also relates to medical uses of the CAR-NK cell or CAR T-cell. The present invention further relates to a CAR-construct, comprising a modified CD8α or CD28 transmembrane domain.

The present disclosure provides an expression cassette comprising from 5' to 3': (i) promoter, (ii) an upstream exon, (iii) an upstream intron, (iv) an alternatively spliced exon comprising a translation initiation sequence, (v) a downstream intron, (vi) a downstream exon, and (vii) a heterologous nucleic acid sequence. In some aspects, the upstream exon, the upstream intron, the alternatively spliced exon, the downstream intron, and the downstream exon, each comprise a portion of a gene selected from the group consisting of a Transient Receptor Potential Cation Channel Subfamily V Member 3 (TRPV3) gene, a Serine / Threonine Kinase 31 (STK31) gene, an Ubiquitin Specific Peptidase 25 (USP25) gene, a IK gene, a Testis Expressed 14, Intercellular Bridge Forming Factor (TEX14) gene, a General Transcription Factor IIB (GTF2B) gene, and a LY6 / PLAUR Domain Containing 3 (LYPD3) gene.

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 specific alternative splicing transcript ARRB1 of B-type inhibin 1 Δexon13 , a specific alternative splicing transcript NCOR2 of nuclear receptor auxiliary inhibin 2 Δexon21 . The application also discloses use of a reagent for detecting the specific alternative splicing transcript ARRB1 of B-type inhibin 1 Δexon13 in preparation of a kit for detecting glioma. The application also discloses use of a reagent for detecting the specific alternative splicing transcript NCOR2 of nuclear receptor auxiliary inhibin 2 Δexon21 in preparation of a kit for detecting glioma. The application also discloses use of a reagent for detecting the specific alternative splicing transcript ARRB1 of B-type inhibin 1 Δexon13 in preparation of a kit for detecting breast cancer. The application proves that ARRB1 Δexon13 and NCOR2 Δexon21 have high specific expression in primary and recurrent glioma, and can be used as one of means for pathological diagnosis or prognosis analysis of glioma.

This invention discloses an antisense oligonucleotide based on tumor splice conversion therapy and its application. Based on the design of antisense oligonucleotides targeting tumor-specific aberrant splicing events (such as PKM, MCL1, and BCLX), the aim is to inhibit tumor growth by inducing splice conversion. Abnormal alternative splicing events are increasingly becoming a marker of cancer and a potential therapeutic target. By designing splice conversion oligonucleotides targeting genes involved in recurrent tumor-specific splicing events (such as PKM, MCL1, and BCLX), this invention successfully regulates aberrant splicing events in tumors, significantly inhibiting tumor growth, and has been validated in various tumor cell lines. Furthermore, this invention further optimizes the design of splice conversion oligonucleotides and explores a bifunctional oligonucleotide strategy to enhance the efficacy of tumor splice conversion therapy.

The present disclosure provides synthetic nucleic acid compositions and methods of use thereof to generate a computational modeling platform to uncover noncoding RNAs that drive disease progression or intrinsic neuroprotection through regulation of splicing.

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.