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Rationale and Objective: Intentional deception (ie, lying) is a complex cognitive act, with important legal, moral, political, and economic implications. Prior studies have identified activation of discrete anterior frontal regions, such as the ventrolateral prefrontal cortex (VLPFC), dorsolateral prefrontal cortex (DLPFC), dorsal medial prefrontal cortex (DMPFC), and anterior cingulate cortex (ACC) during deception. To extend these findings, we used novel real-time functional magnetic resonance imaging (fMRI) technology to simulate a polygraph experience in order to evoke performance anxiety about generating lies, and sought to ascertain the neural correlates of deception., Materials and Methods: In this investigational fMRI study done with a 4-T scanner, we examined the neural correlates of lying in 14 healthy adult volunteers while they performed a modified card version of the Guilty Knowledge Test (GKT), with the understanding that their brain activity was being monitored in real time by the investigators conducting the study. The volunteers were instructed to attempt to generate Lies that would not evoke changes in their brain activity, and were told that their performance and brain responses were being closely monitored., Results: Subjects reported performance anxiety during the task. Deceptive responses were specifically associated with activation of the VLPFC, DLPFC, DMPFC, and superior temporal sulcus., Discussion: These findings suggest the involvement of discrete regions of the frontal cortex during lying, and that the neural substrates responsible for cognitive control of behavior may also be engaged during deception.