Cardiac magnetic resonance imaging feature tracking (CMR-FT) is a novel non-invasive imaging technique and has been validated in clinical applications. However, there are only limited studies in small animal research. We performed biventricular strain assessment using CMR-FT in nexiline knock-out mice at rest and dobutamine stress to evaluate its feasibility and intra-observer reproducibility in preclinic research. Excellent intra-observer reproducibility could be observed for rest and stress left ventricular (LV) strain in general and right ventricular (RV) circumferential strain derived from short-axis views. An only fair reproducibility was observed for RV longitudinal strain under stress conditions derived from long-axis views.