The present disclosure provides methods and compositions for generating populations of auxotrophic cells and populations of differentiated cells and selecting populations of transfected cells using split auxotrophy.

The invention provides methods of determining a therapeutically effective dose of an agent that targets a metabolic pathway based on measured levels of a metabolite in the pathway. The methods, which may include providing the agent in a therapeutically effective dose, are useful for treating disorders, such as cancer, in a subject. The invention also provides methods for assessing the impact of a therapeutic agent on a tumor in a subject by monitoring, in real time, metabolism of a molecule in the tumor, oxygenation of the tumor, or both. The invention further provides devices that determine a therapeutically effective dose of an agent that targets a metabolic pathway based on measured levels of a metabolite in the pathway and notify a subject to administer the dose.

The present invention provides regulatable cell lines with an engineered genome encoding an auxotrophic response system or element, and methods of use thereof. Growth and protein production of the cell lines is controlled by exposure of the cell line to an auxotrophic factor to activate the auxotrophic response system or element. Examples of auxotrophic factors include a nutrient, an enzyme, a moiety that alters pH, a moiety that alters temperature, a non-organic molecule, a non-essential amino acid, an altered concentration of a moiety, and a cellular niche environment. These regulatable cell lines may be used in regenerative medicine and enzyme replacement therapy.

It relates to a Mortierella alpine ATCC32222 uracil auxotroph strain and a construction method thereof. In the present invention, Mortierella alpine ATCC32222 is used as a material and undergoes gene knockout through an Agrobacterium tumefaciens mediated genetic manipulation technology, to obtain the Mortierella alpine uracil auxotroph. The method is of great significance for the basic theoretic researches of the oil producing fungus Mortierella alpine ATCC32222 and product development.

It relates to a Mortierella alpine ATCC32222 uracil auxotroph strain and a construction method thereof. In the present invention, Mortierella alpine ATCC32222 is used as a material and undergoes gene knockout through an Agrobacterium tumefaciens mediated genetic manipulation technology, to obtain the Mortierella alpine uracil auxotroph. The method is of great significance for the basic theoretic researches of the oil producing fungus Mortierella alpine ATCC32222 and product development.