Extracellular vesicles (EVs) are emerging as critical mediators of intercellular communication, carrying bioactive cargoes such as DNA, RNA, and proteins. However, the molecular mechanisms underlying the packaging and release of DNA within these EVs remain unclear. In this study, we identify a distinct class of large, non-lytic cellular fragments, termed Motile Vesicles (MoVes), emitted during active migration of cancer cells. Interestingly, non-transformed epithelial cells do not emit MoVes. We identified MoVes carry double-stranded replicative genomic DNA, histones, and mitochondrial proteins, and exhibit unique proteomic signatures enriched in integrins (ITGA2, ITGA3, ITGB1), Basigin (BSG), and the epithelial-to-mesenchymal transition (EMT) effector FAM3C. Whole-genome sequencing revealed that MoVes DNA mirrors the complete mutational landscape of the parent cancer cells. Notably, migration-inhibitory drugs significantly reduced the biogenesis of MoVes, while the neutral sphingomyelinase 2 (nSMAse2: sEV biogenesis pathway) inhibitor GW4869 had no effect. Moreover, inhibition of TGFβR1 using LY-364947 suppressed cell migration, ultimately preventing DNA emission via MoVes, linking MoVes formation to migration and EMT. In vivo biodistribution of MoVes were most abundant in the liver. Our findings uncover an active, non-apoptotic mechanism for genome-wide DNA release during cancer cells migration, with potential implications for metastasis and extracellular DNA biology.