AbstractTunneling triboelectrification of chemical vapor deposited monolayer MoS₂ has been characterized at nanoscale with contact-mode atomic force microscopy (AFM) and Kelvin force microscopy (KFM). Although charges can be trapped on insulators like SiO₂ by conventional triboelectrification, triboelectric charges tunneling through MoS₂ and localized at the underlying substrate exhibit more than two orders of magnitude longer lifetime. Their polarity and density can be modified by triboelectric process with various bias voltages applied to Pt-coated AFM tips, and the saturated density is almost 30 times higher than the reported result of SiO₂. Thus, the controllable tunneling triboelectric properties of MoS₂ on insulating substrates can provide guidance to build a new class of two-dimensional (2D) MoS₂-based nanoelectronic devices.