Background: Sickle Cell Nephropathy (SCN) is a major cause of morbidity and mortality among patients with Sickle Cell Disease (SCD), arising from chronic hemolysis, oxidative stress, and endothelial dysfunction. Variations in the Endothelial Nitric Oxide Synthase (eNOS) gene may influence Nitric Oxide (NO) bioavailability, potentially affecting renal susceptibility in SCD. Aim: This study aimed to Detect the association of eNOS gene polymorphism (G894T) with nephropathy among Sudanese patients with SCD. Method: A case-control study was conducted between June 2021 and June 2022 at Soba University Hospital and Jafar Ibn Auf Specialized Hospital, Khartoum. Participants included 65 patients with sickle cell nephropathy, 45 nephropathy patients without SCD, and 45 healthy controls. Samples were used for CBC and DNA extraction. ( Hb, PCV, PLTs, WBCs, RBCs count and RBCs indices): Was done by using automated hematology analyzer Genomic DNA was extracted from blood samples and analyzed for G894T polymorphism using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) techniques. Result: The NOS3 (G894T) genotype distributions differed significantly between SCD nephropathy cases and healthy controls (X² = 5.982, p = 0.050). No significant differences were found for G894T or in comparisons between sickle and non-sickle nephropathy groups. Hematological parameters showed no genotype-related variations. Conclusion: The eNOS3 G894T polymorphism showed a significant difference in genotype distribution between cases and normal controls, though logistic regression analysis did not reveal a statistically significant association in genetic models.