Provided herein are improved, cost-effective and environmentally friendly methods of production of recombinant AAV (rAAV) in embryonated avian eggs. Further provided herein is a provides embryonated avian eggs as novel host vehicles for high-yield production of rAAV, including both packaging and propagation. In particular, embryonated chicken eggs provide a novel expression vehicle for AAV of mammalian origin, irrespective of AAV serotype. The disclosed methods may comprise packaging of rAAV in embryonated avian eggs (e.g., chicken eggs) by inoculating an embryonated avian egg with a first nucleic acid vector comprising a transgene and a second nucleic acid vector comprising AAV rep and cap genes, incubating the egg, and isolating rAAV from the egg, wherein the AAV is of non-avian origin. Also provided are methods of purifying and propagating packaged rAAV in embryonated avian eggs or in avian embryonic fibroblasts.

The present invention is transgenic chickens obtained from long-term cultures of avian PGCs and techniques to produce and transgenic birds derived from prolonged PGC cultures. In some embodiments, these PGCs can be transfected with genetic constructs to modify the DNA of the PGC, specifically to introduce a transgene encoding an exogenous protein. When combined with a host avian embryo by known procedures, those modified PGCs are transmitted through the germline to yield transgenic offspring. This invention includes compositions comprising long-term cultures of PGCs and offspring derived from them that are genetically modified. The genetic modifications introduced into PGCs to achieve the gene inactivation may also include, but are not restricted to, random integrations of transgenes into the genome, transgenes inserted into the promoter region of genes, transgenes inserted into repetitive elements in the genome, site specific changes to the genome that are introduced using integrase, site specific changes to the genome introduced by homologous recombination, and conditional mutations introduced into the genome by excising DNA that is flanked by lox sites or other sequences that are substrates for site specific recombination.

A transgenic chicken comprising an inactivated heavy immunoglobulin gene and/or inactivated light chain immunoglobulin gene is provided, as well as cells and targeting vectors for making the same.

The present invention relates to processes for transfecting cells. In particular, the present invention relates to processes for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC).

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

The present invention relates to the production of avian induced pluripotent stem cells from non-pluripotent somatic cells, including embryonic fibroblasts and adult somatic cells. In this method, avian (including quail or chicken) somatic cells are reprogrammed into a state closely resembling embryonic stem cells including the expression of key stem cell markers alkaline phosphatase, etc. by transfecting/transducing the non-stem cells with genes (preferably using a non-integrating vector as otherwise described herein or alternatively an integrating vector, such a lentiviral vector, retroviral vector or inducible lentiviral vector, among others) which express at least nanog, Lin28 and cMyc. In preferred aspects of the invention, the transfected/transduced vectors express nanog, Lig28, cMyc, Oct 4 (POU5F1 or PouV), SOX2 and KLF4. The induced stem cells which are produced contribute to all 3 germ layers, the trophectoderm and in certain aspects, the gonad in chimeric offspring.

The present disclosure provides exogenous polynucleotide cassettes for generating chimeric bird cells and chimeric birds. The polynucleotide cassettes can be used to produce conditionally-lethal phenotype in male bird embryos. In one embodiment, the present disclosure provides methods for destroying male chick embryos in-ovo.

A cage and corresponding method for loading, transporting, and unloading poultry utilize a substantially vertical track attached to a leg or a sidewall to provide stability for a door of the cage throughout the door's entire range of motion. The cage is stackable and together with like cages can form an assembly of cages. The assembled stack of cages allows for several cages to share common components. Thus, the weight of the load during transport is reduced and more livestock can be transported. Shared components can include a roof and even a latching mechanism which prevents the door of each cage from coming open during transport.

The present invention relates to an avian model enabling the amplification of human or animal circulating tumour cells (CTC) and to the use thereof for monitoring and determining the sensitivity of a patient or an animal suffering from cancer to one or more therapeutic agent(s), as well as for screening novel therapeutic agents intended for the treatment of cancer.

A container includes a plurality of panels connected together at fold lines to extend at least partially around an interior space. A plurality of end flaps are each foldably connected to a respective one of the panels. The end flaps are overlapped with one another to enclose an end of the interior space. One of the end flaps is a main bird-feeder flap. A side bird-feeder flap foldably connected to the main bird-feeder flap. The main bird-feeder flap and the side bird-feeder flap are configured to deploy from a closed position cooperating with the plurality of end flaps to enclose the end of the interior space, to an open position with the main bird-feeder flap hinged outward from the interior space to form an opening in the end of the interior space, and with the side bird-feeder flap forming a side of the opening.