21 results about "Trans-splicing" patented technology

The present invention is characterized by nucleic acid trans-splicing molecules (e.g., pre-mRNA trans-splicing molecules (RTMs)) capable of correcting one or more mutations in the ABCA4 or CEP290 genes. Such molecules can be used to treat conditions associated with ABCA4 mutations, such as Stargardt disease (e.g., Stargardt disease 1), and conditions associated with CEP290 mutations, such as Leber congenital amaurosis 10 (LCA 10). The invention also provides methods for using nucleic acid trans-splicing molecules to correct mutations in ABCA4 and CEP290, and for treating conditions associated with mutations in ABCA4 and CEP290, such as Stargardt disease and LCA 10.

A logic-gated protein device in which proximity-gated protein trans-splicing governs formation of an active protein from two otherwise inactive fragments.

Provided is a compact RNA-editing platform engineered from IscB, which has comparable or higher activity than Cas13, but with less or no cytotoxicity. IscB, has intrinsic affinity for complementary single-stranded (ss)DNA and RNA. This activity becomes dominant when its dsDNA binding activity is switched off, through the deletion or mutation of its Target Adjacent Motif domain, resulting in a modified IscB. The resulting R-IscB is comparable or better than Cas13, can efficiently alter splicing outcomes in human cells, and can mediate trans-splicing to correct mutations at the mRNA level. R-IscB also drives efficient A-to-I editing on mRNA when fused to ADAR2 and mediates cleavage-based mRNA knockdown upon HNH engineering. The same approach converts some Cas9s to RNA-targeting tools.

A synthetic nucleic acid expression system for production of a transcript of interest in a predefined cell-state is provided, the system comprising (a) a first nucleic acid sequence comprising a first promoter operably linked to a nucleic acid sequence encoding a first trans-spliceable pre-mRNA sequence comprising at least one exon encoding a 5′ fragment of said transcript of interest and a first RNA sequence required for spliceosome-dependent trans-splicing; and (b) a second nucleic acid sequence comprising a second promoter operably linked to a nucleic acid sequence encoding a second trans-spliceable pre-mRNA sequence comprising at least one exon encoding a 3′ fragment of said transcript of interest and a second RNA sequence required for spliceosome-dependent trans-splicing; wherein said first promoter and said second promoter are different and each one is specifically regulated by said predefined cell-state.

An exon editor construct is provided comprising a binding domain that binds to a target intron of MeCP2 pre-mRNA, a hemi-intron, and a coding domain comprising one or more MeCP2 exons. A method for expressing functional MeCP2 in target cells is also provided, comprising contacting target cells with the exon editor construct provided herein.

The present disclosure provides methods and compositions for RNA trans-splicing. Also disclosed are methods of treating a condition, disease, or disorder in a subject using the disclosed methods and compositions.

This invention discloses a method and its application for predicting the trans-splicing activity of fragmented inpeptides. The method takes the amino acid sequence of the fragmented inpeptide as input, extracts 1280-dimensional residue-level sequence embedding features using the ESM-2 protein language model, and uses AlphaFold2 for high-precision three-dimensional structure prediction to obtain the spatial coordinates of the residues. A graph structure is constructed with residues as nodes and residue pairs with a Cα atomic distance of less than or equal to 8 Å as edges. Node features include sequence embedding and chain affiliation information, while edge features include Cα distance and cross-chain connection identifiers. The graph structure is input into the DeepIntein deep learning model based on a graph attention network, and the splicing activity prediction probability is output after multi-layer graph attention convolution, global pooling, and fully connected layers. The prediction throughput of this invention is more than three orders of magnitude higher than that of the traditional Western blotting method, breaking through the limitations of homology sequence exploration space.

Disclosed herein are compositions for and methods of modulating RNA trans-splicing and methods of using disclosed compositions in methods treating and / or preventing a genetic disease or disorder with the mRNA generated via trans-splicing.

The present invention provides pre-mRNA trans-splicing molecules (RTMs) that are useful for correcting mutations in the OPA1 gene. Also provided are methods of using the RTMs as gene therapy (e.g., ex vivo and in vivo gene therapy) for the treatment or prevention of diseases or disorders associated with OPA1 mutations.

The invention relates to a trans-splicing system. The trans-splicing system comprises a U2 type or U12 type splicing donor / splicing acceptor pair. In a preferred embodiment, the orthogonality of the trans-splicing system is further improved by adding an identification module. The trans-splicing system can be advantageously used for constructing a logic line, expressing nucleic acid molecules of interest, detecting cell types, detecting target RNA, constructing a virus genetic barrier, performing targeted therapy and the like.

Disclosed are a multi-vector expression system encoding precursor mRNA molecules and precursor mRNA trans-splicing molecules (PTM), and a cultured eukaryotic recombinant host cell comprising the system. Furthermore, disclosed are a method for recombinantly expressing one or more target proteins in a cultured eukaryotic host cell line, and a method for screening a eukaryotic host cell recombinantly expressing a target protein in vitro, which employ the multi-vector expression system of the present application.

The invention discloses a recombinant expression and in-vitro splicing integrated preparation method and application of spider silk protein, and belongs to the technical field of biology. Coding genes of NT, Rep or CT regions of spider silk protein from at least two different spider species are fused, a fusion gene sequence is optimized by pichia pastoris codon and then introduced into an Intein tag, and the spider silk protein is obtained. Dividing the full-length protein into an N-terminal fragment and a C-terminal fragment; respectively cloning the N-terminal fragment and the Intein-N structural domain as well as the C-terminal fragment and the Intein-C structural domain into a pichia pastoris secreting type expression vector pPICZalpha; respectively electrically transforming the two obtained vectors into pichia pastoris SMD1168 for induced expression, and collecting supernatant; sequentially carrying out Ni-NTA affinity chromatography and ion exchange chromatography on the supernatant to obtain a purified N-terminal protein fragment and a purified C-terminal protein fragment; mixing the spider silk protein and the spider silk protein, forming full-length spider silk protein through Intein-mediated trans-splicing, and purifying through size exclusion chromatography. The method provided by the invention can realize high-efficiency and high-quality production of spider silk protein.

The present invention provides compositions, systems, and methods for using ribozyme-mediated cis-cleavage and trans-splicing or cis-ligation of RNA molecules to express a target protein or fusion protein.

The present invention features nucleic acid trans-splicing molecules (e.g., pre-mRNA trans-splicing molecules (RTMs)) capable of correcting one or more mutations in the ABCA4 gene or the CEP290 gene. Such molecules are useful in the treatment of disorders associated with mutations in ABCA4, such as Stargardt Disease (e.g., Stargardt Disease 1) and disorders associated with a mutation in CEP290, such as Leber congenital amourosis 10 (LCA 10). Also provided by the invention described herein are methods of using the nucleic acid trans-splicing molecules for correcting mutations in ABCA4 and CEP290 and for treating disorders associated with mutations in ABCA4 and CEP290, such as Stargardt Disease and LCA 10.

Nucleic acids, vectors, compositions, systems, and methods for expressing a structural maintenance of chromosomes hinge domain containing 1 (SMCHD1) polypeptide to epigenetically silence double homeobox 4 (DUX4) for the treatment of a disease or disorder associated with DUX4 are provided. DUX4 regulates gene expression and plays a role in development, muscular dystrophy (including, but not limited to, facioscapulohumeral dystrophy (FSHD)), a cancer, or Bosma arhinia microphthalmia syndrome (BAMS). The disclosure describes a split intein-mediated protein trans-splicing approach that was utilized to express SMCHD1 to downregulate or inhibit DUX4 expression.

This disclosure relates to compositions, systems, and methods associated with repair RNA sequences, and related systems for trans-splicing target nucleic acids using repair RNA sequences. Various compositions and systems are described herein, including compositions comprising a repair RNA sequence containing one or more exons and / or introns, a splicing donor, and / or a splicing acceptor, wherein the repRNA is adapted for trans-splicing. The compositions, systems, and methods described herein are suitable for targeting one or more Usher syndrome-related genes and for treating Usher syndrome.

A nucleic acid trans-splicing molecule is provided that can replace an exon in a targeted mammalian ocular gene carrying a defect or mutation causing an ocular disease with an exon having the naturally-occurring sequence without the defect or mutation. A method of treating an ocular disease, e.g., Stargardt's Disease, caused by a defect or mutation in a target gene, e.g., ABCA4 comprising: administering to the ocular cells of a subject having an ocular disease a composition comprising a recombinant AAV comprising a nucleic acid trans-splicing molecule as described above.

The present invention provides compositions, systems and methods for using ribozyme-mediated cis-cleavage and trans-splicing of RNA molecules to express a functional SCN5A, and methods of use thereof for treating Brugada Syndrome and other diseases associated with SCN5A loss of function.

Compositions and methods for efficient construction of circular RNAs are described. In particular, a method of trans-splicing involving multiple segments of a Group I intron or derivatives thereof is described for efficient production of circular RNAs substantially free of dimeric or concatemeric byproducts. Compositions related to the method are also described.

Provided herein are nucleic acid trans-splicing molecules (e.g., pre-mRNA trans-splicing molecules (RTMs); RNA exon editing molecules) capable of correcting mutations in the ABCA4 gene. Such molecules are useful in the treatment of disorders such as ABCA4-associated retinal dystrophies (e.g., Stargardt Disease or cone-rod dystrophy). Also described herein are methods of using the nucleic acid trans-splicing molecules described herein to correct mutations in ABCA4, thereby treating disorders associated with mutations in ABCA4 and use of the nucleic acid trans-splicing molecules described herein for treating disorders associated with mutations in ABCA4 and in the preparation of medicaments for the treatment of disorders associated with mutations in ABCA4.

Described herein are compositions systems and methods for promoting trans-splicing. In some examples, the system may comprise a nucleic acid molecule. The nucleic acid molecule may encode an exonic sequence. The system may further comprise a tethering fusion protein. The tethering fusion protein may promote an association of the exonic sequence and a target RNA or portion thereof.