The present invention relates to the field of fibrosis and inflammation and more particularly to the use of ADAM12 (A Disintegrin and Metalloproteinase 12) inhibitors to prevent or treat inflammation-induced fibrosis. The present invention also relates to the use of ADAM12 as a marker for inflammation-induced fibrosis and to the ablation of ADAM12 expressing cells as therapeutic approach to interfere with the development of pro-fibrotic cells.

Provided are a Moloney murine leukemia virus-based self-inactivating vector and applications thereof. The self-inactivating vector comprises: 5LTR, a target expression gene or multiple cloning sites used for inserting the target expression gene, 3LTR, and a polyadenylated nucleic acid fragment, where the nucleic acid fragment between the Pvu II enzyme cutting site and the Sac I enzyme cutting site of the U3 region of 5LTR is replaced by a CMV enhancer and a CMV promoter connected to the CMV enhancer, the nucleic acid fragment between the Pvu II enzyme cutting site and the Sac I enzyme cutting site of the U3 region of 3LTR is deleted, and the polyadenylated nucleic acid fragment is located at the 3-terminus of 3LTR. The self-inactivating vector, serving as a vector for a gene therapy, provides increased safeness and a high virus titer, and allows the high-efficiency expression of a target gene.

The invention relates to the production and use of Cas-encoding sequences and vectors comprising these. Aspects of the invention provide products, vectors, delivery vehicles, uses and methods for producing Cas-encoding sequences in bacterial or archaeal cells.

A eukaryotic replicative pUC-free minicircle expression vector is provided. The eukaryotic replicative pUC-free minicircle expression vector includes a pUC-free eukaryotic region sequence encoding a transgene of interest and comprising 5 and 3 ends and a ii) pUC-free spacer region of less than 500 basepairs in length linking the 5 and 3 ends of the eukaryotic region sequences and comprising a bacterial R6K replication origin having at least 95% sequence identity to SEQ ID NO: 11 and SEQ ID NO: 12 and a RNA selectable marker, the RNA selectable marker being an RNA-IN regulating RNA-OUT functional variant having at least 95% sequence identity to SEQ ID NO: 20 or SEQ ID NO: 22.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from the group consisting of inverted repeat sequence, direct repeat sequence, homopolymeric repeat sequence, eukaryotic origin of replication or eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 by from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Poi I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.

The invention provides an expression and secretion system, and methods of using the same, for the expression and secretion of one fusion protein in prokaryotic cells and a second fusion protein in eukaryotic cells. Also provided herein are nucleic acid molecules, vectors and host cells comprising such vectors and nucleic acid molecules.

A eukaryotic replicative pUC-free minicircle expression vector is provided. The eukaryotic replicative pUC-free minicircle expression vector includes a pUC-free eukaryotic region sequence encoding a transgene of interest and comprising 5 and 3 ends and ii) a pUC-free spacer region of less than 500 basepairs in length linking the 5 and 3 ends of the eukaryotic region sequences and comprising a bacterial R6K replication origin having at least 95% sequence identity to SEQ ID NO: 11 or SEQ ID NO: 12 and a RNA selectable marker, the RNA selectable marker being an RNA-IN regulating RNA-OUT functional variant having at least 95% sequence identity to SEQ ID NO:20 or SEQ ID NO:22.

The present invention relates to a polynucleotide comprising at least one promoter and an S/MAR element, wherein said S/MAR element is located downstream of said promoter in the 3 UTR of the transcription unit and wherein the said S/MAR element is flanked by a 5 splice donor site and a 3 splice acceptor site; the present invention further relates to a composition comprising said polynucleotide, and to the polynucleotide for use in medicine and for use in treating genetic disease.

The present description relates to genetic tools and methods to facilitate transformation and genetic engineering of industrially-useful yeast/fungal species, such as Issatchenkia orientalis, for which a robust set of genetic tools such as stably inherited and maintained plasmids and functional control sequences is presently lacking. Thus, the present description relates to autonomously replicating sequences (ARSs), RNA polymerase II/III promoters, RNA polymerase II/III terminators, expression cassettes, and vectors comprising same are described herein, as well as uses and methods relating to same, which are functional in I. orientalis.

The present invention relates to the field of self-replicating non-integrative episomal vertebrate expression vectors useful for in gene therapy, ex vivo cell therapy, stem cell therapy, and more particularly, for improving the expression of vector encoded antigens or therapeutic genes. Such recombinant DNA molecules are useful in biotechnology, transgenic organisms, gene therapy, stem cell therapy, therapeutic vaccination, agriculture and DNA vaccines. More specifically, relates to a polynucleotide comprising at least one promoter and an S/MAR element, wherein said S/MAR element is located downstream of said promoter and wherein the nucleic acid sequence of said S/MAR element (S/MAR sequence) comprises at least 3 sequence motifs ATTA (SEQ ID NO:1) per 100 nucleotides over a stretch of at most 200 nucleotides; the present invention further relates to a composition and to a host cell comprising said polynucleotide, and to the polynucleotide for use in medicine and for use in treating genetic disease. The present invention also relates to a kit and to a device comprising said polynucleotide, and to methods and uses related to the polynucleotide.