Provided herein are compositions and methods to make and use engineered cells, for the purpose of increasing the cell density of a culture comprising metazoan cells and for the production of a cultured edible product.

Methods of inserting genes into defined locations in the chromosomal DNA of cultured mammalian cell lines which are subject to gene amplification are disclosed. In particular, sequences of interest (e.g., genes encoding biotherapeutic proteins) are inserted proximal to selectable genes in amplifiable loci, and the transformed cells are subjected to selection to induce co-amplification of the selectable gene and the sequence of interest. The invention also relates to meganucleases, vectors and engineered cell lines necessary for performing the methods, to cell lines resulting from the application of the methods, and use of the cell lines to produce protein products of interest.

Methods of inserting genes into defined locations in the chromosomal DNA of cultured mammalian cell lines which are subject to gene amplification are disclosed. In particular, sequences of interest (e.g., genes encoding biotherapeutic proteins) are inserted proximal to selectable genes in amplifiable loci, and the transformed cells are subjected to selection to induce co-amplification of the selectable gene and the sequence of interest. The invention also relates to meganucleases, vectors and engineered cell lines necessary for performing the methods, to cell lines resulting from the application of the methods, and use of the cell lines to produce protein products of interest.

The present invention provides for a mechanism to reduce glycerol production and increase nitrogen utilization and ethanol production of recombinant microorganisms. One aspect of this invention relates to strains of S. cerevisiae with reduced glycerol productivity that get a kinetic benefit from higher nitrogen concentration without sacrificing ethanol yield. A second aspect of the invention relates to metabolic modifications resulting in altered transport and/or intracellular metabolism of nitrogen sources present in corn mash.

Disclosed herein are methods of increasing nitrogen fixation in a non-leguminous plant. The methods can comprise exposing the plant to a plurality of bacteria. Each member of the plurality comprises one or more genetic variations introduced into one or more genes or non-coding polynucleotides of the bacteria's nitrogen fixation or assimilation genetic regulatory network, such that the bacteria are capable of fixing atmospheric nitrogen in the presence of exogenous nitrogen. The bacteria are not intergeneric microorganisms. Additionally, the bacteria, in planta, produce 1% or more of the fixed nitrogen in the plant.

The present disclosure concerns a recombinant bacterial host cell capable of providing a nitrogen source to a yeast during fermentation to make ethanol. The recombinant bacterial host cell is capable of converting a biomass into ethanol. The recombinant bacterial host cell has at least one first genetic modification. The at least one genetic modifications confers to the recombinant bacterial host cell the ability to increase, when compared to a corresponding control bacterial cell lacking the at least one first genetic modification, the proteolytic activity associated with the recombinant bacterial host cell. The at least one genetic modification also confers the recombinant bacterial host cell the ability to provide a nitrogen source to a yeast capable of converting the biomass into ethanol, wherein the nitrogen source comprises a peptide, an amino acid and/or ammonia.

Provided herein are compositions and methods to make and use engineered cells, for the purpose of increasing the cell density of a culture comprising metazoan cells and for the production of a cultured edible product.

A reagent for glutamine synthetase reaction comprising a chelating agent and glutamine synthetase, and a reagent for quantification of ammonia comprising a chelating agent, ATP, glutamic acid, glutamine synthetase, glucose, an oxidized NAD compound, ADP-dependent hexokinase, and glucose-6-phosphate dehydrogenase, are provided.

The present invention provides methods and compositions for stable genetic modification of cultured mammalian cells. The genetic modifications can be used to produce cultured mammalian cells for therapeutic or diagnostic purposes.

A glycosylation-modified erythropoietin, containing a glycan structure incorporated into an N-glycosylation site, wherein the glycan structure contains FA4G4L2S4.