Background Glucose-6-phosphate dehydrogenase deficiency, known as G6PD deficiency, is a common hematological disease in the Eastern Province. The presence of Glucose-6-phosphate dehydrogenase enzyme in the erythrocyte is crucial, as it aids in the protection of RBCs by preventing cellular damage. It was found that more than 400 million people in the world lack this enzyme, making it the most common enzyme deficiency worldwide. Because of the high prevalence of the disease in the world and the paucity of research in Saudi Arabia about G6PD deficiency, the idea of examining and assessing the awareness and knowledge of parents who have children affected with G6PD emerged. Objective This study aimed to evaluate parental knowledge regarding G6PD deficiency, identify the common and spreadmisconceptions regarding the disease, provide general insightfor physicians about parental knowledge, and propose strategies to educate parents about G6PD deficiency. Methods Thiscross-sectional study was conducted from September 2022 to May 2023 on 459 individuals from the Eastern Province, Saudi Arabia. Using aquestionnaire, parents with variable education levels and incomes participated. The questionnaire was available in bothArabic and English. The study aimed to gather comprehensive data regarding parental awareness and knowledge of G6PD deficiency, it focused on evaluating levels of awareness, knowledge, and misinformation of the participants. Data were analyzed by using Statistical Package for the Social Sciences, ver22 (IBM Corp., Armonk, NY). The chi-square test was applied to check any association between demographics and level of knowledge (Good and Poor). Multiple logistic regression analysis was performed on significant demographic variables and an odds ratio with a 95% confidence interval was calculated. P < 0.05 was considered statistically significant. Results The study included a total of 459 participants, 309 (67.3%) were females and 150 (32.7%) were males. Regarding the demographics, the majority of the participants were <40 years old (61.9%), Saudi (99.3%), married (98.5%), having bachelor's/diploma level or above (Master/PhD) (76.9%), having inherited disorder (44.2%), and having a family history of chronic diseases (82.6%). Furthermore, regarding the knowledge of the disease, the majority of the participants who recognized the disease (82.1%)were familiar with the term fava bean anemia rather than it being called G6PD deficiency anemia. On the other hand, 73.2%of the participants were unable to recognize the disease and had never heard of G6PD deficiency anemia. In addition, the majority of participants had deficient information regarding medication triggers (61.4%), whether the gender of a person is linked to G6PD (77.5%), and whether both parents must be carriers to have a child with G6PD deficiency anemia (50.9%). Female respondents proved to be more knowledgeable regarding this topic as 57.9%of them showed good knowledge as compared to (44.7%) of the male subjects. Conclusion There are discrepancies in the level of awareness among research participants. Our result indicates the need for educational interventions regarding the nomenclature, medication triggers, inheritance mode and its relation to gender, and the symptoms of the disease and its severity. Therefore, it is advised to spread awareness in the eastern province through brochures, medical campaigns, and by healthcare professionals in different medical organizations.