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.

Disclosed herein are cells expressing DUX4 including stem cells, differentiated cells thereof, primary T cells, and chimeric antigen receptor T cells, as well as related methods of their use and generation. In some embodiments, the cells disclosed herein do not express one or more MHCI and/or MHC II human leukocyte antigens. In some embodiments, such cells possess immune evasion properties.

Provided herein, in some embodiments, are methods and compositions for gene delivery. Provided herein is a technology for co-delivering to a cell (e.g., in vivo or ex vivo) enzymes capable of rearranging nucleic acid, such as site-specific recombinases, to directly assemble (e.g., covalently join) nucleic acid segments of, for example, a gene of interest.

Methods of producing hepatocyte lineage cells are provided. The method can include transfecting a cell with one or more expression vectors. For example, a cell can be transfected with a first expression vector containing a first gene that encodes CCAAT/enhancer binding protein alpha (CEBPA), a second expression vector containing a second gene that encodes CCAAT/enhancer binding protein beta (CEBPB), a third expression vector containing a third gene that encodes forkhead box A1 (FOXA1), and a fourth expression vector containing a fourth gene that encodes forkhead box A3 (FOXA3). The method can include culturing the transfected cell obtained in a growth environment. The transfected cell can be cultured in Williams' E medium, ReproFF (feeder-free media maintaining pluripotency) medium, or both. Transfected and/or hepatocyte lineage cells obtained by a method of the present invention are also provided.

The disclosure features compositions and methods for the treatment of sensorineural hearing loss and auditory neuropathy, particularly forms of the disease that are associated with a mutation in otoferlin (OTOF), by way of OTOF gene therapy. The disclosure provides a variety of compositions that include a first nucleic acid vector that contains a polynucleotide encoding an N-terminal portion of an OTOF isoform 5 protein and a second nucleic acid vector that contains a polynucleotide encoding a C-terminal portion of an OTOF isoform 5 protein. These vectors can be used to increase the expression of OTOF in a subject, such as a human subject suffering from sensorineural hearing loss.

Provided herein are compositions that include at least two different nucleic acid vectors, where each of the at least two different vectors includes a coding sequence that encodes a different portion of an otoferlin protein, and the use of these compositions to treat hearing loss in a subject.

The present invention is directed to a single input multiplex decision expression system, and a method of using same, such as for expressing a reporter protein.

Novel therapeutic immunotherapy compositions comprising at least two vectors, each vector encoding a functional CAR, whereby the combination of vectors results in the expression of two or more non-identical binding domains, wherein each vector encoded binding domain(s) are covalently linked to a transmembrane domain and one or more non-identical intracellular signaling motifs are provided herein as well as are methods of use of same in a patient-specific immunotherapy that can be used to treat cancers and other diseases and conditions.

Provided herein are methods and compositions for expressing Otoferlin, e.g., utilizing adeno-associated viral (AAV) particles. Further provided herein are compositions of AAV particles comprising one or more polynucleotides encoding Otoferlin are codon optimized for expression in human cells. Such methods and compositions may be useful for treatment of diseases and disorders such as Deafness, Autosomal Recessive 9 (DFNB9). Also provided are kits comprising such compositions.

The disclosure features compositions and methods for the treatment of sensorineural hearing loss and auditory neuropathy, particularly forms of the disease that are associated with mutations in otoferlin (OTOF), by way of OTOF gene therapy. The disclosure provides a variety of compositions that include a first nucleic acid vector that contains a polynucleotide encoding an N-terminal portion of an OTOF protein and a second nucleic acid vector that contains a polynucleotide encoding a C-terminal portion of an OTOF protein. These vectors can be used to increase the expression of OTOF in a subject, such as a human subject suffering from sensorineural hearing loss.