Guilt is a central moral emotion due to its inherent link to norm violations, thereby affecting both individuals and society. Furthermore, the nature and specificity of guilt is still debated in psychology and philosophy, particularly with regard to the differential involvement of self-referential representations in guilt relative to shame. Here, using functional magnetic resonance imaging (fMRI) in healthy volunteers, we identified specific brain regions associated with guilt by comparison with the 2 most closely related emotions, shame and sadness. To induce high emotional intensity, we used an autobiographical memory paradigm where participants relived during fMRI scanning situations from their own past that were associated with strong feelings of guilt, shame, or sadness. Compared with the control emotions, guilt episodes specifically recruited a region of right orbitofrontal cortex, which was also highly correlated with individual propensity to experience guilt (Trait Guilt). Guilt-specific activity was also observed in the paracingulate dorsomedial prefrontal cortex, a critical "Theory of Mind" region, which overlapped with brain areas of self-referential processing identified in an independent task. These results provide new insights on the unique nature of guilt as a "self-conscious" moral emotion and the neural bases of antisocial disorders characterized by impaired guilt processing.