Dual recombinase-mediated endothelial cell-specific lineage tracing and ablation.

The precise ablation of specific cell lineages is crucial for functional studies in vivo. Conventional methods, like the Cre-dependent iDTR system, are constrained by the off-target effects and variable efficiency of single-recombinase approaches. Here, we present a novel Cdh5-RL-DTRGFP mouse model that requires both Dre and Cre recombinases to activate diphtheria toxin receptor (DTR) and GFP expression specifically in endothelial cells. This dual-recombinase logic ensures tight control over transgene expression. We demonstrate that diphtheria toxin administration in recombined mice leads to efficient endothelial cell ablation, resulting in severe vascular leakage, rapid organ failure, and mortality. The Cdh5-RL-DTRGFP line thus provides a robust and precise platform for the genetic dissection of endothelial cell function in physiological and pathological contexts.