A method of preparing a recombinant adeno-associated virus (rAAV) including: (1) preparing a shuttle plasmid and a corresponding recombinant bacmid including a baculovirus genome, where the shuttle plasmid includes at least an rAAV gene of interest flanked by inverted terminal repeats (ITR-GOI) integrated with a heterologous functional gene fragment, and the recombinant bacmid includes an expression cassette of functional protein components necessary for assembly of the rAAV; (2) integrating the rAAV ITR-GOI and the expression cassette of functional protein components by using the shuttle plasmid and the recombinant bacmid, to yield a recombinant bacmid including a recombinant baculovirus genome; and (3) transfecting, with the recombinant bacmid, a host cell line.

In one aspect, the embodiments disclosed herein relate to the production of fully-deleted adenovirus-based gen delivery vectors packaged without the use of an adenoviral helper virus, and more particularly in their use in the transfer of genes and the expression of proteins, vaccine development, and cell engineering. In another aspect, the production of adenoviral vectors deleted of all adenoviral genes is described that carry genes of interest with detrimental or toxic activities to eukaryotic cells.

Provided herein are adeno-associated virus (AAV) compositions that can restore phenylalanine hydroxylase (PAH) gene function in cell. Also provided are methods of use of the AAV compositions, and packaging systems for making the AAV compositions.

The embodiments disclosed herein relate to the construction of fully-deleted Adenovirus-based gene delivery vectors packaged without helper Adenovirus, and more particularly to their use in gene therapy for gene and protein expression, vaccine development, and immunosuppressive therapy for allogeneic transplantation. In an embodiment, a method for propagating an adenoviral vector includes (a) providing an Adenovirus packaging cell line; (b) transfecting a fully-deleted Adenoviral vector construct into the cell line; and optionally (c) transfecting a packaging construct into the cell line, wherein the fully-deleted Adenoviral vector construct and optionally the packaging construct can transfect the Adenovirus packaging cell line resulting in the encapsidation of a fully-deleted Adenoviral vector independent of helper Adenovirus. In an embodiment, a target cell is transduced with the encapsidated fully-deleted Adenoviral vector for treating a condition, disease or a disorder.

A fast and accurate three-plasmid oncolytic adenovirus recombinant packaging system Ad5MixPlus and an application thereof are provided. The system is composed of three adenovirus recombinant plasmids. The core technology of the system is that two sets of different site recombination sequences are skillfully loaded on a first 5-type adenovirus right arm backbone plasmid large vector, then two small shuttle plasmids respectively provide a right arm-modified Hexon/E3/Fiber sequence and an E1a expression cassette controlled by a left arm tumor-specific promoter, and the difficulties and obstacles to the modification of the adenovirus backbone large vector are overcome. After two rounds of site-specific recombination, the ideal oncolytic adenovirus is packaged accurately and quickly.

During lung fibrosis, activation of fibroblasts to myofibroblasts, is accompanied by the increased assembly of extracellular matrix proteins fibronectin and collagen. Activated myofibroblasts are key drivers of fibrosis; and increased deposition of the assembled matrix proteins, preserves the myofibroblast state. These changes contribute to enhanced tissue stiffness, collagenous deposition known as fibroblastic foci in lung and therefore, established pulmonary fibrosis. The present invention identifies a novel function of the 40 amino acid SOCS domain peptide (SEQ ID NO:1), in degrading the pathologic fibronectin and collagen matrix associated with fibrosis, markedly reducing the myofibroblast protein α-SMA and reducing collagen deposits in the fibrotic lung, leading to fibrosis reversal. Use of the SOCS domain peptide is a new approach to treat lung fibrosis as it restricts extracellular matrix assembly and reverses myofibroblasts to fibroblasts.

The present invention relates to a cell line for producing an adenovirus having a limited autoreplication capability, and a method of preparing the same, and more particularly, to a cell line for producing an adenovirus having no autoreplication capability by expressing at least one selected from an adenoviral E1 protein and an E1A or E1B protein, and a method of preparing the same. Also, the present invention relates to the use of the cell line expressing at least one selected from the adenoviral E1 protein and the E1A or E1B protein.

The invention provides adeno-associated virus (AAV) replication (Rep) sequences. In one embodiment, the invention provides nucleotide sequences encoding a chimeric protein, wherein the encoded chimeric protein contains a wild type AAV Rep inhibitory amino acid sequence, and wherein the nucleotide sequences contain a scrambled and/or deoptimized polynucleotide sequence encoding the wild type AAV Rep inhibitory amino acid sequence. The invention provides vectors, cells, and viruses containing the invention's sequences. Also provided are methods for detecting portions of the AAV Rep inhibitory amino acid sequence, which reduce replication and/or infection and/or productive infection by viruses. The invention's compositions and methods are useful for site-specific integration and/or expression of heterologous sequences by recombinant adeno-associated virus (rAAV) vectors and by rAAV virus particles, such as hybrid viruses (e.g., Ad-AAV) comprising such vectors. The invention's compositions and methods find application in, for example, gene therapy and/or vaccines.

Provided herein are adeno-associated virus (AAV) compositions that can restore phenylalanine hydroxylase (PAH) gene function in cell. Also provided are methods of use of the AAV compositions, and packaging systems for making the AAV compositions.

A method of preparing a recombinant adeno-associated virus (rAAV) including: (1) preparing a shuttle plasmid and a corresponding recombinant bacmid including a baculovirus genome, where the shuttle plasmid includes at least an rAAV gene of interest flanked by inverted terminal repeats (ITR-GOI) integrated with a heterologous functional gene fragment, and the recombinant bacmid includes an expression cassette of functional protein components necessary for assembly of the rAAV; (2) integrating the rAAV ITR-GOI and the expression cassette of functional protein components by using the shuttle plasmid and the recombinant bacmid, to yield a recombinant bacmid including a recombinant baculovirus genome; and (3) transfecting, with the recombinant bacmid, a host cell line.