The present invention relates to a recombinant vector, characterized by including a cytotoxic T-lymphocyte-associated protein-4 (CTLA-4)-targeting trans-splicing ribozyme expression cassette for delivery of chimeric antigen receptor, wherein the expression cassette includes: (i) a CTLA-4-targeting trans-splicing ribozyme; and (ii) a polynucleotide encoding a chimeric antigen receptor ligated to the 3 exon of the ribozyme. The present invention also relates to a transformed cell into which the recombinant vector is introduced, a ribozyme expressed from the recombinant vector, a retrovirus expressing the ribozyme, and a T cell treated with the retrovirus. Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating cancers, in which the pharmaceutical composition includes the recombinant vector, the transformed cell, the ribozyme, the retrovirus, the T cell, or a combination thereof; and a method for treating cancers, in which the method includes administering, to an individual in need thereof, the recombinant vector, the transformed cell, the ribozyme, the retrovirus, the T cell, or a combination thereof. The recombinant vector of the present invention and the ribozyme expressed therefrom become a gene-cell therapy which inhibits CTLA-4 on T cells which has been an obstacle in conventional anti-cancer therapies and, at the same time, enables anti-cancer treatment, thereby allowing more effective anti-cancer effects to be anticipated. Such a gene-cell therapy results in decreased toxicity in normal tissues and thus exhibits increased effects in both therapeutic efficacy and safety, which enables it to be widely utilized in the field of gene therapy in the future.

A self-circularization RNA construct that can be expressed in a DNA vector and simultaneously circularized through a self-targeting and splicing reaction to form a circRNA is disclosed. The circRNA can consist only of a gene of interest which can be a coding, non-coding, or a combination thereof. The gene of interest has the advantage of being able to rapidly express a peptide or protein. The formed circRNA has a circular structure and has a stable and high half-life because 5 and 3 ends are not exposed. Accordingly, functional RNA such as miRNA, anti-miRNA, siRNA, shRNA, aptamer, functional RNA for gene/RNA editing, ADAR (adenosine deaminase acting on the RNA)-recruiting RNA, mRNA vaccine, mRNA therapeutic agent, vaccine adjuvant, and CAR-T mRNA can be produced as a stable circRNA in cells.

The present invention relates to a recombinant vector, characterized by including a cytotoxic T-lymphocyte-associated protein-4 (CTLA-4)-targeting trans-splicing ribozyme expression cassette for delivery of chimeric antigen receptor, wherein the expression cassette includes: (i) a CTLA-4-targeting trans-splicing ribozyme; and (ii) a polynucleotide encoding a chimeric antigen receptor ligated to the 3 exon of the ribozyme. The present invention also relates to a transformed cell into which the recombinant vector is introduced, a ribozyme expressed from the recombinant vector, a retrovirus expressing the ribozyme, and a T cell treated with the retrovirus. Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating cancers, in which the pharmaceutical composition includes the recombinant vector, the transformed cell, the ribozyme, the retrovirus, the T cell, or a combination thereof; and a method for treating cancers, in which the method includes administering, to an individual in need thereof, the recombinant vector, the transformed cell, the ribozyme, the retrovirus, the T cell, or a combination thereof. The recombinant vector of the present invention and the ribozyme expressed therefrom become a gene-cell therapy which inhibits CTLA-4 on T cells which has been an obstacle in conventional anti-cancer therapies and, at the same time, enables anti-cancer treatment, thereby allowing more effective anti-cancer effects to be anticipated. Such a gene-cell therapy results in decreased toxicity in normal tissues and thus exhibits increased effects in both therapeutic efficacy and safety, which enables it to be widely utilized in the field of gene therapy in the future.

The present invention relates to a recombination vector, a transformation cell into which the recombinant vector is introduced, a ribozyme expressed from the recombination vector, a prophylactic or therapeutic composition for liver cancer comprising the recombination vector and the ribozyme, and a therapeutic method for liver cancer using the composition, said recombination vector comprising: a tissue-specific promoter; and a ribozyme-target gene expression cassette comprising a trans-splicing ribozyme targeting a cancer-specific gene and a target gene connected to the 3 exon of the ribozyme, wherein a splicing donor/splicing acceptor sequence (SD/SA sequence) is connected to the 5 end of the ribozyme-target gene expression cassette, woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) is connected to the 3 end of the ribozyme-target gene expression cassette, and a nucleic acid sequence recognizing a micro RNA-122a (microRNA-122a, miR-122a) is further connected to the 3 end of the WPRE.

Provided are methods for editing RNA by introducing a deaminase-recruiting RNA in a host cell for deamination of an adenosine in a target RNA. Further provided are deaminase-recruiting RNAs used in the RNA editing methods and compositions and kits comprising the same.

The present invention relates to a recombination vector, a transformation cell into which the recombinant vector is introduced, a ribozyme expressed from the recombination vector, a prophylactic or therapeutic composition for liver cancer comprising the recombination vector and the ribozyme, and a therapeutic method for liver cancer using the composition, said recombination vector comprising: a tissue-specific promoter; and a ribozyme-target gene expression cassette comprising a trans-splicing ribozyme targeting a cancer-specific gene and a target gene connected to the 3 exon of the ribozyme, wherein a splicing donor/splicing acceptor sequence (SD/SA sequence) is connected to the 5 end of the ribozyme-target gene expression cassette, woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) is connected to the 3 end of the ribozyme-target gene expression cassette, and a nucleic acid sequence recognizing a micro RNA-122a (microRNA-122a, miR-122a) is further connected to the 3 end of the WPRE.

An intronic, self-splicing riboswitch is configured for enzyme-product specificity by introducing an appropriate aptamer. This then provides a sensing-expression construct, whereby the presence of an enzyme product in the cell triggers self-splicing of the intron sequence to restore the reading frame of the reporter gene and as such to drive expression of the gene product. The sensing construct expresses a protein which marks the cell or permits its growth or survival in or on an otherwise selective media. In this way, introduction or the presence of such product sensing-reporter constructs in cells can be harnessed to provide a multi-parallel rapid screening of cells or libraries for desirable enzyme variants.

A method of preparing a circular RNA includes transcribing a vector to form a precursor RNA, in which the vector includes the following elements operably connected to each other and arranged in the following sequence: a) a 5 element, b) a 3 Group I self-splicing intron fragment containing a 3 splice site dinucleotide, c) none or an element containing an internal ribosome entry site (IRES) and a protein coding region or an element containing a noncoding region, d) a 5 Group I self-splicing intron fragment containing a 5 splice site dinucleotide, and e) a 3 element, in which the 5 element and the 3 element form a stable structure with a Gibbs free energy (G) from 190 kcal/mol to 9.0 kcal/mol, provided that the stable structure is not a duplex with at least 95% base pairing between the 5 element and the 3 element, in which the 3 Group I self-splicing intron fragment and the 5 Group I self-splicing intron fragment form a self-cleaving and self-ligating RNA molecule, thereby generating circular RNA.

The present invention relates to the technical field of nucleic acids, and in particular, to a method for precisely preparing a circular RNA with an Anabaena intron self-cleaving ribozyme. By redesigning a target nucleic acid sequence of a linear RNA, selecting a site suitable for precise cleavage by the intron self-cleaving ribozyme, and removing exon sequences from the Anabaena intron self-cleaving ribozyme, precise cyclization of RNAs after in vitro transcription is achieved. The method provided by the present invention solves the problem in the prior art that linear RNAs cannot be precisely cyclized during the process of generating circular RNAs in vitro, and provides an ideal in vitro preparation method for circular RNA applications.

Described is a unique class of antiviral molecule that can be applied to control and eliminate HIV infection in patients using myeloablation therapies and replenishment with transformed bone marrow stem cells programmed to express the antiviral molecule. These anti-viral molecules target the HIV genome in a highly conserved domain, and when expressed in cells prior to infection will cause the cell to die upon infection with HIV. Cell death insures no proliferation of new virus. Reconstituting the immune system with cells expressing these antivirals prevents re-establishment of HIV infection from reservoirs in the re-established lymphocyte and macrophage populations. Over time, reservoirs will be depleted entirely, effectively eliminating the virus. In effect, this new type of antiviral can be used to cure HIV infections.