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Mental health research emphasizes the importance of fostering character strengths to promote well-being. However, whether the strengths supposedly developed by these interventions actually mediate their effects on well-being is still unknown. The included studies were RCTs of strength-based interventions on well-being. Among 83,908 reviewed abstracts, five strengths met the strict inclusion criteria to be meta-analyzed. We included 114 RCTs with a total of 20,853 participants. A One-Stage Meta-Analytic Structural Equation Modeling approach was used to combine studies and analyze the mediation models. Perspective-based interventions showed improvements in perspective. Kindness-based interventions and gratitude-based interventions demonstrated moderate associations with their respective strengths. Humor and hope-based interventions did not significantly enhance their respective trained strengths. Significant mediation effects on well-being were found for all strengths, except hope. The robust pattern of mediations aligns with eudaemonic theories, emphasizing the importance of virtue-driven actions in fostering psychological well-being.