The presently disclosed subject matter relates to targeted integration (TI) host cells suitable for the expression of recombinant proteins wherein those TI host cells have been subjected to supertransfection resulting in the random integration (RI) of exogenous nucleic acids encodes into their genome, as well as methods of producing and using said supertransfected TI host cells.

A transgenic animal having a genome including a humanized immunoglobulin locus for securing the diversity of an antibody repertoire and a method of producing the same are disclosed. The transgenic non-human animal has two alleles of a humanized immunoglobulin gene, wherein the two alleles are hetero-alleles.

Provided herein are compositions and methods for high throughout cloning of fused bipartite immunoreceptor polynucleotides encoding cognate pairs of bipartite immunoreceptors. Also provided herein are various applications of the fused bipartite immunoreceptor polynucleotides, expression vectors containing the fused bipartite immunoreceptor polynucleotides, or cells containing the fused bipartite immunoreceptor polynucleotides or expression vectors.

Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a rodent (e.g., a mouse), wherein the non-human animals rearrange human immunoglobulin light chain gene segments in the context of heavy chain constant regions and express immunoglobulin-like molecules comprising human immunoglobulin light chain variable domains fused to heavy chain constant domains that are cognate with human immunoglobulin light chain variable domains fused to light chain constant domains.

A combined system comprising two separate components, wherein the first component is a tag-exchange donor vector (TEDV) encoding a first cell surface tag (CST) exon flanked by a 3′ intron fragment, and a gene of interest (GOI) in the antisense orientation, and the second component is an engineered cell containing within its genome a tag-exchange receiver site (TERS), encoding a second CST exon adjoined by a full intron sequence to an exon encoding a transmembrane domain, and also encoding a reporter gene in the antisense orientation, wherein paired recombinase mediated cassette exchange (RMCE) elements are included in the TEDV and TERS such that execution of RMCE between the TEDV and TERS results in exchange of the reporter element for the GOI encoded by the TEDV, and exchange of the first CST exon for the second CST exon, such that the derivative engineered cell now expresses the first CST and GOI, in place of the second CST and the reporter gene.

Methods, compositions and non-human animals and parts thereof are for improving germ line transmission of genetic modifications. The methods and compositions are for producing non-human embryos with a disrupted or disruptable fertility gene. The embryos can be used as hosts for the development of donor pluripotent cells, including genetically modified donor pluripotent cells, into germ cells and gametes. Additional methods and compositions are for producing from such embryos chimeric non-human animals with a disrupted fertility gene and for breeding the chimeric non-human animals with cognate non-human animals that comprise a fertility gene that lacks a disruption to produce non-human animals having substantially all gametes and/or germ cells derived from the donor pluripotent cells. Non-human gametes, germ cells, embryos and animals can be used in the subject methods.

Methods for integrating exogenous sequences in genomic loci, wherein the integration is stable and the exogenous sequence can function predictably and reliably.

Described herein are methods of inserting nucleic acid sequences into host cells. Also described herein are genetically stable host cells comprising inserted nucleic acid sequences and methods of using such host cells in the generation of proteins.

Mice, tissues, cells, and genetic material are provided that comprise a humanized heavy chain immunoglobulin locus, a humanized light chain locus that expresses a universal light chain, and a gene encoding an ADAM6 or ortholog or homolog or functional fragment thereof. Mice are provided that express humanized heavy chains comprising human variable domains, and that express humanized light chains comprising human variable domains wherein the light chains are derived from no more than one, or no more than two, light chain V and J or rearranged V/J sequences. Fertile male mice that express antibodies with universal light chains and humanized heavy chains are provided. Methods and compositions for making bispecific binding proteins are provided.

The invention relates to an adeno-associated virus (AAV) vector producer cell comprising nucleic acid sequences encoding AAV rep and cap genes, helper virus genes, and a DNA genome of the AAV vector; the AAV rep gene comprising an intron, the intron comprising a transcription termination 5 sequence with a first recombination site located upstream and a second recombination site located downstream of the transcription termination sequence; and the nucleic acid sequences all integrated together at a single locus within the AAV vector producer cell genome. The invention also relates to methods for producing the AAV vector producer cell lines.