Figure 1.
Swapping out CENP-A at the centromere with an inducible gene replacement strategy. Gene editing is first used to add an auxin-inducible degron (AID) sequence to endogenous CENP-A, which undergoes rapid TIR1 E3 ubiquitin ligase-dependent degradation in the presence of the plant hormone auxin. Degraded CENP-AAID is then rescued by a doxycycline-inducible gene encoding a CENP-A/histone H3 carboxy-tail chimera (CENP-AC-H3) that does not support kinetochore assembly specifically on the Y centromere, thereby producing Y chromosome-selective segregation errors in the subsequent mitosis.