Background/Objectives: Extraction and replacement of hopeless teeth is a common practice in dentistry. Tooth autotransplantation (TA) offers several advantages as a viable and biological treatment option. However, its utilization in Saudi Arabia appears limited. Understanding dentists’ knowledge and attitudes toward TA is crucial for promoting its adoption in clinical practice. Therefore, the aim of this study is to assess the knowledge and attitudes of dentists in Saudi Arabia toward tooth autotransplantation. Methods: A cross-sectional study was conducted using a web-based questionnaire distributed to dentists across Saudi Arabia. The questionnaire included 19 questions assessing demographic characteristics, knowledge of TA, and attitudes toward its clinical application. Data were analyzed using SPSS v23 with chi-square and Cramér’s V tests to explore associations between variables (p < 0.05). Results: 253 dentists participated in this study. All participants had heard of TA, while only 26.5% reported moderate-to-high familiarity. Female dentists showed significantly greater interest in adopting TA than males (p = 0.038, Cramér’s V = 0.183). Specialists expressed higher familiarity than general dentists and underscored the importance of clinical guidelines, evidence-based outcomes, confidence in their capacity to engage in a TA team, the total number of required appointments, and malpractice concerns (p < 0.05) in their decision-making process. Nevertheless, general dentists demonstrated significantly more interest in implementing TA (p = 0.025, Cramér’s V = 0.192). Participants with more than 5 years of clinical dental experience were significantly more familiar with TA (p = 0.015, Cramér’s V = 0.204) and were more influenced by appointment numbers in decision-making (p = 0.012, Cramér’s V = 0.225). Conclusions: The study reveals limited familiarity but notable interest among dentists in Saudi Arabia toward TA. Addressing educational gaps by integrating TA training into dental curricula and offering clinical exposure opportunities to students, along with providing evidence-based clinical guidelines, and improving access to advanced imaging technologies, may enhance the adoption of TA as a viable treatment modality for tooth replacement.