Neurobiological studies of tobacco/nicotine use examining genetic, molecular, functional, and behavioral correlates have improved our understanding of nicotine/tobacco dependence and have informed treatment. Recent work extending previously established findings and reporting novel methodologies and discoveries in preclinical and human studies are reviewed.Recent work in preclinical models has focused on the differential roles of nicotinic receptor subtypes and nicotine's effects on neural systems beyond cortico-striatal dopaminergic pathways, and utilizing advanced methodologies such as pharmacogenetics, optogenetics and rodent fMRI to identify targets for treatment. Likewise, human neuroimaging studies have identified molecular and functional dynamic shifts associated with tobacco/nicotine use that further inform treatment.Nicotine/tobacco use is associated with widespread neural adaptations that are persistent and function to maintain addiction. The continued identification of genetic, molecular, neural, and behavioral endophenotypes related to nicotine/tobacco use, dependence, and addiction will facilitate the development and delivery of personalized treatment.