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 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.

Replicating recombinant adenovirus vectors derived from human adenovirus serotype 26 or human adenovirus serotype 35 are described. The replicating recombinant adenovirus vectors have attenuated replicative capacity as compared to that of the corresponding wild-type adenovirus. They can be used for stable expression of heterologous genes in vivo. Also described are compositions and methods of using these recombinant adenovirus vectors to induce an immune response in a subject, and vaccinate a subject against an immunogenic human immunodeficiency virus (HIV) infection.

The present invention provides recombinant nucleic acid expression cassetie and helper dependent adenovirus, where the expression cassettes utilize a miRNA based system for controlling expression of nucleases in helper dependent adenoviral viral producer cells, thus permitting production and use for in in vivo gene editing in CD34+ cells.

An african swine fever virus vaccine includes five groups of antigens in total, and each group is respectively obtained by constructing recombinant adenovirus vectors co-expressing four antigen genes of african swine fever virus, and packaged by 293TD37 cells. The four antigenic genes of African swine fever virus in each group are 1, P72, B602L, P30 and P54; 2, CP129Rubiqutin, MGF5L6L, CP312R, and MGF110-4L; 3, L8Lubiqutin, I215L, I73RHBsAgHBsAg and E146L; 4, EP402R, EP153R, I177L, and K205Rubiqutin; 5, F317L, A151R, P34, and pp62. The construction of the recombinant adenovirus vector for co-expression of four antigen genes of the african swine fever virus mainly includes: knocking out E1, E3, E2a and E4 genes of the adenovirus vector by CRISPR/cas9 technology, constructing an ORF6/7 expression frame of E4 in an E2a region, and constructing shuttle plasmids in E1 and E4 regions for appropriately expressing four antigen genes, thereby obtaining a completely new adenovirus vector.

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.

Embodiments provide systems and methods for transfecting cells to produce a viral vector or other selected virus. One embodiment of a method for transfecting cells to produce a selected virus includes providing a first solution comprising plasmids or other extrachromosomal DNA encoding the virus and a second solution comprising a transfection agent; the two solutions kept separate. The two solutions are then mixed to produce a transfection solution (TS) to be delivered to a cell culture medium (CCM) for transfection of cells in the medium with the plasmid DNA encoding the virus wherein the mixing is initiated responsive to a trigger event. The TS is then incubated to form transfection complexes. Subsequently, the TS is delivered to the CCM to transfect the cells in the medium wherein a viral production parameter resulting from transfection is optimized by initiating mixing of the solutions responsive to the trigger event.

The present application relates to the technical field of biology, and specifically, to an exogenous gene controllable expression virus packaging vector and a packaging method. The virus packaging vector has two ITR fragments and a gene expression cassette inserted between the two ITR fragments. The gene expression cassette comprises a promoter, a repressor operator, and an optional target gene that are connected in sequence. When there is a repressor, the repressor operator can repress the expression of a downstream target gene thereof.

The present disclosure relates to a method for producing a recombinant adenovirus vector.

The present invention provides a method of releasing viral vectors from cells producing those viral vectors by contacting the cells with a photosensitising agent which is then irradiated to disrupt the plasma membrane of the cells to release the viral vectors which may be collected and/or purified. The product of such methods as well as kits and apparatuses for performing the methods are also provided.