Background: Permanent neonatal diabetes (PND) is a monogenic form of diabetes resulting from mutations in a number of different genes encoding proteins that play a key role in the normal function of the pancreatic beta-cell. A correct genetic diagnosis can affect treatment and clinical outcome. Mutations in the insulin gene (INS) itself have been identified as a cause of neonatal diabetes. This study aimed to investigate the genetic variations in the coding region and intronic boundaries of INS gene and their genotype phenotype correlation in a group of Egyptian PNDM infants with onset in the first 12 months of age. Methods: We screened exons 2 and 3 with intronic boundaries of INS gene by direct gene sequencing in 30 PND patients (14 males and 16 females) and in 20 healthy control subjects to verify the resulting single nucleotide polymorphisms (SNPs). A detailed clinical phenotyping of the patients was carried out to specify the diabetes features in those found with an INS variant. Results: We identified five variants (four SNPs and one silent mutation), c.187+11T>C, c.-17-6T>A, c.*22A>C, c.*9C>T and c.36G>A (p.A12A) with allelic frequency of 96.7%, 80%, 75%, 5% and 1.7%, respectively. All showed no statistically significance difference compared to the controls except for c.*22A>C. Conclusion: Genetic screening for the INS gene did not reveal any evident role in diagnosis of PNDM among the studied group of Egyptian children.