Stoichiometric Cr2Se3 single crystals are particular layer-structured antiferromagnets, which possess a noncollinear spin configuration, weak ferromagnetic moments, moderate magnetoresistance (MR ∼14.3%), and poor metallic conductivity below the antiferromagnetic phase transition. Here, we report an interesting >16 000% colossal magnetoresistance (CMR) effect in Ti (1.5 atomic percent) lightly doped Cr2Se3 single crystals. Such a CMR is approximately 1143 times larger than that of the stoichiometric Cr2Se3 crystals and is rarely observed in layered antiferromagnets and is attributed to the frustrated spin configuration. Moreover, the Ti doping not only dramatically changes the electronic conductivity of the Cr2Se3 crystal from a bad metal to a semiconductor with a gap of ∼15 meV but also induces a change in the magnetic anisotropy of the Cr2Se3 crystal from strong out-of-plane to weak in-plane. Further, magnetotransport measurements reveal that the low-field MR scales with the square of the reduced magnetization, which is a signature of CMR materials. The layered Ti:Cr2Se3 with the CMR effect could be used as two-dimensional (2D) heterostructure building blocks to provide colossal negative MR in spintronic devices.