Background: despite the worldwide recognition of the importance of pre-screening education of parents, the current body of evidence suggests that parents often receive little information about neonatal screening (NS) and may even be unaware about the screening of their baby. Aim of the work: this study aimed to assess the attitudes and knowledge of the Saudi women towards the NS program and their psychological impact. Methods: we performed a cross-sectional study on mothers in the Eastern Region of the Kingdom of Saudi Arabia. A modified version of the Maternal Attitudes and Knowledge about Newborn Screening Survey was used to assess the attitudes and knowledge of the women towards the NS program. Results: we retrieved 388 surveys. Twenty-five percent of the women acquired their knowledge about NS programs from the internet. Eighty-two percent of the women had healthy children and 42% of them think that the best time to know about NS programs is at the time of screening itself. Our analysis showed a significant association between the educational level and knowledge about the best time to learn about neonatal screening (p=0.0001). Almost half of the women who had one child stated that the screening should take place 2-3 months before the baby is born (p=0.018). The child health status was also associated with the knowledge that if the baby's newborn test is abnormal I might have something wrong with my DNA (p=0.015). It was also associated with knowing that NS will test for common diseases that run in families like diabetes, asthma and heart disease (p=0.02). Conclusion: Saudi women had a positive attitude, but with little knowledge towards the NS program. Further studies are needed to assess the predictors of different levels of knowledge.