Efficient conditional mutation of the vertebrate CENP-C gene

We have used gene targeting in the DT40 cell line to create a cell linewhich expresses a fusion between CENP-C and a mouse steroid receptor andwhich behaves as a conditional loss of function mutant of CENP-C. Underrestrictive conditions these cells arrest at the metaphase/anaphasejunction and after a delay of approximately 2.5 h die by apoptosis. Theseresults indicate that CENP-C is either necessary for anaphase chromosomemovement or for mediating a signal which triggers centromere functionduring anaphase. Our approach is simple and applicable to a wide range ofproteins with general cell autonomous functions in vertebrates.