Background There is an ongoing debate on the off-hour effect on endovascular treatment (EVT) for acute large vessel occlusion (LVO). Aims This meta-analysis aimed to compare time metrics and clinical outcomes of acute LVO patients who presented/were treated during off-hour with those during working hours. Summary of review Structured searches on the PubMed, Embase, Web of Science, and Cochrane Library databases were conducted through February 23rd, 2021. The primary outcomes were onset to door, door to imaging, door to puncture, puncture to recanalization, procedural time, successful recanalization, symptomatic intracerebral hemorrhage (SICH), mortality in hospital, good prognosis (90-day modified Rankin Scale [mRS] score 0-2), and 90-day mortality. The secondary outcomes were imaging to puncture, onset to puncture, onset to recanalization, door to recanalization time, mRS 0-2 at discharge, and consecutive 90-day mRS score. The odds ratio (OR) and weighted mean difference (WMD) with 95% confidence interval (CI) of the outcomes were calculated using random-effect models. Heterogenicity and publication bias were analyzed. Subgroup and sensitivity analyses were conducted as appropriate. Nineteen studies published between 2014 and 2021 with a total of 14185 patients were eligible for quantitative synthesis. Patients in the off-hour group were significantly younger than those in the on-hour group and with comparable stroke severity and intravenous thrombolysis rate. The off-hour group had longer onset to door (WMD [95%CI], 12.83 [1.84-23.82] min), door to puncture (WMD [95%CI], 11.45 [5.93-16.97] min), imaging to puncture (WMD [95%CI], 10.39 [4.61-16.17] min), onset to puncture (WMD [95%CI], 25.30 [13.11-37.50] min), onset to recanalization (WMD [95%CI], 25.16 [10.28-40.04] min), and door to recanalization (WMD [95%CI], 18.02 [10.01-26.03] min) time. Significantly lower successful recanalization rate (OR [95%CI], 0.85 [0.76-0.95]; P=0.004; I2=0%) was detected in the off-hour group. No significant difference was noted regarding SICH and prognosis. But a trend towards lower OR of good prognosis was witnessed in the off-hour group (OR [95%CI], 0.92 [0.84-1.01]; P=0.084; I2=0%). Conclusions Patients who presented/were treated during off-hour were associated with excessive delays before the initiation of EVT, lower successful reperfusion rate, and a trend towards worse prognosis when compared with working hours. Optimizing the workflows of EVT during off-hour is needed.