Tuberous sclerosis complex (TSC) is a genetic disorder caused by mutations inTSC1orTSC2. Patients frequently have epilepsy, autism spectrum disorder, and/or intellectual disability, as well as other systemic manifestations. In this study, we differentiated human induced pluripotent stem cells (iPSCs) from a female patient with TSC with one or two mutations inTSC2into neurons using induced expression of NGN2 to examine neuronal dysregulation associated with the neurological symptoms in TSC. Using this method, neuronal differentiation was comparable between the three genotypes of iPSCs. We observed thatTSC2^+/−neurons show mTOR complex 1 (mTORC1) hyperactivation and associated increased cell body size and process outgrowth, as well as exacerbation of the abnormalities by loss of the second allele ofTSC2inTSC2^−/−neurons. Interestingly, iPSC-derived neurons with either a single or biallelic mutation inTSC2demonstrated hypersynchrony and downregulation of FMRP targets. However, only neurons with biallelic mutations ofTSC2demonstrated hyperactivity and transcriptional dysregulation observed in cortical tubers. These data demonstrate that loss of one allele ofTSC2is sufficient to cause some morphological and physiological changes in human neurons but that biallelic mutations inTSC2are necessary to induce gene expression dysregulation present in cortical tubers. Finally, we found that treatment of iPSC-derived neurons with rapamycin reduced neuronal activity and partially reversed gene expression abnormalities, demonstrating that mTOR dysregulation contributes to both phenotypes. Therefore, biallelic mutations inTSC2and associated molecular dysfunction, including mTOR hyperactivation, may play a role in the development of cortical tubers.SIGNIFICANCE STATEMENTIn this study, we examined neurons derived from induced pluripotent stem cells with two, one, or no functionalTSC2(tuberous sclerosis complex 2) alleles and found that loss of one or both alleles ofTSC2results in mTORC1 hyperactivation and specific neuronal abnormalities. However, only biallelic mutations inTSC2resulted in elevated neuronal activity and upregulation of cell adhesion genes that is also observed in cortical tubers. These data suggest that loss of heterozygosity ofTSC1orTSC2may play an important role in the development of cortical tubers, and potentially epilepsy, in patients with TSC.