Nitric oxide (NO) regulates blood pressure (BP) by binding the reduced heme iron (Fe2+) in soluble guanylyl cyclase (sGC) and relaxing vascular smooth muscle cells (SMC). We previously showed that sGC heme iron reduction (Fe3+ → Fe2+) is modulated by cytochrome b5 reductase 3 (CYB5R3). However, the in vivo role of SMC CYB5R3 in BP regulation remains elusive. Here, we generated conditional smooth muscle cell-specific Cyb5r3 knockout mice (SMC CYB5R3 KO) to test if SMC CYB5R3 loss impacts systemic BP in normotension and hypertension via regulation of sGC redox state. SMC CYB5R3 KO mice exhibited a 5.84 mmHg increase in BP and impaired acetylcholine-induced vasodilation in mesenteric arteries compared to controls. To drive sGC oxidation and elevate BP, we infused mice with angiotensin-II. We found SMC CYB5R3 KO mice exhibited a 14.75 mmHg BP increase and mesenteric arteries had diminished NO-dependent vasodilation, but increased responsiveness to sGC heme-independent activator BAY 58-2667 over controls. Furthermore, acute injection of BAY 58-2667 in angiotensin-II treated SMC CYB5R3 KO mice showed greater BP reduction compared to controls. Together, these data provide the first in vivo evidence that SMC CYB5R3 is a sGC heme reductase in resistance arteries and provides resilience against systemic hypertension development.