Background: Neural tube defects are one of the most common birth defects. The most frequent forms of neural tube defects are anencephaly and myelomeningocele (Spina bifida) which occurs due to incomplete closure of neural tube at the cranial and caudal end respectively. Infants with anencephaly die in utero or are still born. Among all VANGL1 has been shown to be in close relation with the disease. The present study aimed to investigate the mutation in VANGL1 gene among Pakistani population having myelomeningocele. Material & Methods: The study was conducted among fifty diagnosed cases of myelomeningocele with age group from 0-10 years. Several anatomical patterns were analyzed like site and size of the cyst, by prenatally on ultrasound. While taking history several parameters were considered like folic acid consumption by the mothers and the consequences of the disease on siblings. Blood samples of the patients were drawn and PCR was performed for VANGL1 gene mutations were identified by DNA sequencing. Result: In this study we came across the variation of the disease which was more prevalent in males. Most common site was the lumbar region, showing the size of the cyst. On further investigation, it revealed that about 96% of mothers did not take folic acid during their pregnancy and 1.4% of patients showed positive family history having myelomeningocele. During our study we came across the rare mutation of VANGL1 in five patients. The mutations identified in VANGL1 gene which were at different substitution position of amino acid i.e. valine 239 with Phenylalanine. Conclusion: With this study we have finally concluded that in future further investigations and diagnosis for neural tube disorders should be carried out. In other cases although history revealed myelomeningocele, mutation should be identified. So keeping this point in consideration, other gene mutation should be done and more research of same area based should be carried out.