Abstract: Disclosed are constructs and methods for expressing DNAs of interest in particular in non-primate eukaryotic host cells that display advantages with regard quantity and quality of expression including high stability of expression and, if appropriate, transport of the expression product out of the cell.

Abstract: A transcriptomic analysis of genes consistently upregulated in high producer clones were each evaluated for their ability to increase the production of a protein of interest. The products of these genes (metabolism influencing products (MIP)), such as actin, Erp27, Erp57, Foxa1, PPAR, Ca3, and Tagap, could be sub-categorized into different functional categories such as signaling, protein folding, cytoskeleton organization and cell survival.

Abstract: Disclosed are cells that have stably integrated into their genomes exogenous nucleic acid sequences, such as transgenes, within or proximal to the integration site of a sequence comprising at least part of an endogenous retrovirus (ERV) or a LTR-retrotransposon (LTR-RT), or instead of a sequence encompassing an ERV or a LTR-RT that is part or was part of the genome of the cell, as well as method of producing and using such cells. Advantageously, a high level and/or stable production of the transgene expression product(s) can be achieved. Transgene integration and expression may be furthered by modulating the DNA repair pathways of the cell, e.g., by transiently expressing a gene encoding a protein that forms part of a DNA repair pathway during transgene integration.

Abstract: The present disclosure provides production systems and host cells to produce collagen 7 compositions comprising recombinant collagen 7 and/or functional variants thereof. The host cells are genetically engineered to stably express rCol7 and functional variants thereof. The collagen 7 composition can be used to restore collagen 7 levels in a subject in need, and for preventing, preventing the progression of, alleviating, and delaying the on-set of a skin condition, e.g., skin wound associated with dystrophic epidermolysis bullosa (DEB).

Abstract: Disclosed herein are methods of genome alteration, in particular genome editing in eukaryotic cells (e.g., mammalian cells), preferably, but not exclusively the integration of exogenous nucleic acids into the genome of a cell or a population of cells. Such methods include the modulation of cell cycle phases via external conditions such as physical separation, temperature, exposure to certain substances such as cell cycle modulators. Genome alteration is also effected via the use of enzymes such as nucleases and nickases and/or the modulation of DNA repair pathways.

Abstract: Disclosed are eukaryotic expression systems and methods for the selection of mammalian cell lines that produce proteins of interest, such as therapeutic proteins. The systems and methods allow for a simple and fast selection of cells mediating high levels of recombinant protein production. The systems and methods decrease the efforts and time needed to bring a new therapeutic protein to the patients, and also lower the cost of the therapeutic protein by increasing the productivity of cells in a bioreactor.

Abstract: Disclosed are constructs and methods for expressing DNAs of interest in particular in non-primate eukaryotic host cells that display advantages with regard quantity and quality of expression including high stability of expression and, if appropriate, transport of the expression product out of the cell.

Abstract: Disclosed are methods and eukaryotic host cells for transgene expression. The cells may be treated and/or modified to increase homologous recombination (HR), decrease non homologous end joining (NHEJ) and/or to enhance a HR/NHEJ ratio in said cell. Such cells can be transfected with vectors comprising the transgene, which advantageously integrates into the genome of the cell to form a concatemeric structure which may comprise more than 200 transgene copies. Certain expression enhancing elements such as MARs are advantageously provided to further enhance and/or facilitate transgene expression. Disclosed is also a recombinant eukaryotic host cell, in particular a non-primate host cell, comprising a transgenic sequence encoding a protein and/or a RNA, in particular a primate protein and/or RNA, involved in translocation across the ER membrane and/or secretion across the cytoplasmic membrane.