Abstract Background : Tenecteplase is a tissue plasminogen activator with higher fibrin specificity compared with Alteplase. Accumulating data suggests that intravenous Tenecteplase 0.25mg/kg is non-inferior to Alteplase 0.9mg/kg for acute ischemic stroke. We describe our 10-months experience. Methods : At our MRI-based, urban comprehensive stroke center, we switched the intravenous thrombolytic agent for acute ischemic stroke to Tenecteplase 0.25mg/kg on March 23, 2021. Until January 31, 2022, 62 stroke patients were treated with Tenecteplase. We compared clinical and safety outcomes of Tenecteplase-treated patients with 94 Alteplase-treated patients. Results : During the study period, nine (15%) patients with unknown stroke onset were thrombolyzed with MRI screening. Nineteen (35%) patients underwent subsequent thrombectomy. When compared with Alteplase-treated patients, there was no difference with Tenecteplase-treated patients in 90-day functional outcome, death, symptomatic intracranial hemorrhage, or angioedema. Conclusions : The use of Tenecteplase for stroke thrombolysis was feasible with comparable safety and functional outcomes compared to Alteplase, even when Tenecteplase was administered based on MRI screening to stroke patients with unknown onset.