Striatal cholinergic interneurons (SCIN) are key modulators of motor learning, cognitive flexibility, and strategy selection. Our previous work showed that SCIN ablation did not impair motor skills but altered spatial navigation, hindering learning and increasing perseveration. Yet, how SCIN contribute to different phases of motor learning and perseverative behaviors remains unclear. To elucidate this, adult heterozygous Chat-Cre mice (males/females) were injected with a viral vector to express an inhibitory DREADD (hSyn-DIO-hM4D-Gi-mCherry) in the SCIN. Three weeks later, vehicle (control) and CNO-treated mice were tested in an accelerating rotarod task (4–40 rpm) across 5 consecutive days (5–7 trials/day). CNO was injected only on Day 1, while vehicle was administered throughout. CNO-treated mice showed enhanced performance compared to controls on Day 1, suggesting that SCIN inhibition at this stage may influence strategy selection. To further examine perseverance, adult heterozygous Chat-IDTR mice (males/females) received diphtheria toxin (DT) to ablate SCIN, followed weeks later by corpus callosum lesions which disconnected the orbitofrontal cortex (OFC) from the striatum. DT and control mice were tested before and after surgery in marble burying, hole board, and Y-maze tasks. DT mice displayed perseverative behaviors, which might be alleviated after OFC-striatal disconnection. In summary, our findings suggest SCIN involvement is more multifaceted than initially thought.