Several prospective multigenerational studies have shown that crime runs in the family, while empirical research on the biological causes of crime has also established that low heart rate is related to antisocial behavior. This study examines whether the intergenerational transmission of crime is moderated or mediated by a low heart rate of the son. Prospectively collected conviction data on 794 men from three consecutive generations of the Dutch Transfive dataset is used. Heart rates were measured around age 18, during the medical examination prior to the mandatory military service in the Dutch army. All analyses were conducted separately for violent and non-violent crime. Both paternal violence and low heart rate levels are associated with increased violent offending. Intergenerational transmission of violence was only found among families in which the son had a low heart rate, although the degree of transmission did not differ significantly from families in which the son had a high heart rate. No support was found for a mediating influence of low heart rates of criminals’ offspring on the intergenerational transmission of crime and violence. The results from this study underline the importance to focus on the interaction between biological risk factors and psychosocial risk factors for criminal behavior.