Background: Maxillary canines are the second most frequently impacted teeth after third molars, with a prevalence of approximately 2% in the general population. The etiology of canine impaction is multifactorial, involving both environmental and genetic factors. Among the candidate genes associated with tooth development, PAX9 plays a crucial role in odontogenesis and tooth bud positioning. Genetic variations in PAX9 have been linked to dental anomalies such as agenesis and impaction; however, evidence regarding their association with maxillary canine impaction remains limited, particularly in the Indian population. Aim: This study aimed to evaluate the association between PAX9 gene polymorphism (SNP rs4904210) and maxillary canine impaction in an Indian population sample. Materials and Methods: A case-control study was conducted on 60 subjects (30 cases with impacted maxillary canines and 30 controls with normally erupted canines) aged 10–40 years. Venous blood samples were collected, and DNA was isolated using standard protocols. The PAX9 gene was amplified using Polymerase Chain Reaction (PCR), followed by Restriction Fragment Length Polymorphism (RFLP) analysis with the enzyme HpaII for genotyping. Statistical analyses, including Hardy-Weinberg equilibrium and chi-square tests, were performed to determine the association between genotypes and the occurrence of canine impaction. Results: The study population showed no significant differences in age or gender distribution between cases and controls (p > 0.05). Among the cases, unilateral impaction was more common (73.3%) than bilateral (26.7%). Both case and control groups were in Hardy-Weinberg equilibrium. No statistically significant differences in genotype or allele frequencies of PAX9 (rs4904210) were observed between the two groups, indicating no genetic association with maxillary canine impaction. Conclusion: The present study found no significant association between PAX9 polymorphism (rs4904210) and maxillary canine impaction in the Indian population studied. Although PAX9 plays a critical role in tooth development, this specific SNP does not appear to influence canine impaction risk. Future studies with larger, ethnically diverse samples and broader genetic analysis are warranted to better understand the multifactorial etiology of maxillary canine impaction.