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Positive behavior support (PBS) has been widely emerged as an effective framework to support positive behaviors and address challenging behaviors. To effectively implement PBS, professional development is essential for teachers. However, research indicates that one-time workshops have limited effectiveness. Thus, the purpose of this study was to employ an online three-tier teacher training (3TTT) model to implement a PBS curriculum, and to explore its effects on the knowledge improvement among six general education teachers. The three-tier model consisted of asynchronous self-study, synchronous group workshop, and one-on-one coaching. Using a single-subject changing conditions design, quantitative data were visually analyzed, and qualitative data as well as participant feedback were integrated to support the findings of the study. The findings revealed that: 1) Two out of six teachers met the response criteria at Tier 2, 2) knowledge improved for five out of six teachers, with two teachers meeting the response criteria, 3) all five teachers agreed that applying behavioral skills training was beneficial for learning and practice, and four teachers had a positive attitude towards the PBS curriculum, 4) four out of five teachers agreed that the online 3TTT model aligned with their learning styles and preferences, and provided different levels of support, and 5) four out of five teachers believed that PBS was helpful in managing student behavior problems, expressed willingness to use it in the future, and demonstrated confidence in preventing potential behavior problems. This study represents the first in Taiwan utilizing the three-tier training model. The findings highlighted the utility of three-tier model, the importance of diverse learning resources, and the necessity of behavior skills training. Future research may consider expanding the scale of the 3TTT, strengthening behavior skill training through practical skill practice, extending PBS curriculum and exploring changes in teacher behavior.