Inducible site-directed mutagenesis through conditional gene rescue

A method for producing an inducible site-directed mutant cell capable of conditional gene rescue, comprising

• a) introducing in a target cell a mutated allele of a gene to be mutated by using a suitable mutagenesis technique,
• b) introducing in said target cell a rescue allele of said gene that can be conditionally inactivated, and
• c) optionally, cultivating said target cell under conditions that allow for a selection of cells that contain both the mutated allele and the rescue allele of said gene,
• wherein said mutation in said mutated allele of said gene interferes with survival and/or causes an adverse phenotype.

A method for the production of an inducible site-directed non-human mutant-organism comprising a cell capable of conditional gene rescue, comprising

• a) generating an inducible site-directed mutant cell by a method comprising
• i) introducing in a target cell a mutated allele of a gene to be mutated by using a suitable mutagenesis technique,
• ii) introducing in said target cell a rescue allele of said gene that can be conditionally inactivated, and
• iii) optionally, cultivating said target cell under conditions that allow for a selection of cells that contain both the mutated allele and the rescue allele of said gene,
• wherein said mutation in said mutated allele of said gene interferes with survival and/or causes an adverse phenotype;
• and
• b) generating a non-human mutant organism comprising said mutant cell.

An inducible site-directed non-human mutant-organism, produced according to a method comprising

• a) generating an inducible site-directed mutant cell by a method comprising
• i) introducing in a target cell a mutated allele of a gene to be mutated by using a suitable mutagenesis technique,
• ii) introducing in said target cell a rescue allele of said gene that can be conditionally inactivated, and
• iii) optionally, cultivating said target cell under conditions that allow for a selection of cells that contain both the mutated allele and the rescue allele of said gene,
• wherein said mutation in said mutated allele of said gene interferes with survival and/or causes an adverse phenotype;
• and
• b) generating a non-human mutant organism comprising said mutant cell.