The V2O5 self-assembled nanosheets (SANsV2O5) were synthesized by an additive-free ultrasonic method with subsequent thermal decomposition. The loose nanostructured V2O5 is multi-layer structure stacked by 4∼6 layers with a thickness of 200∼300nm and each layer is composed by abundant nanoparticles in order. Due to this unique layered structure, the SANs V2O5 exhibits remarkable rate performance and excellent cycling stability. Specifically, it delivers reversible discharge capacities of 157.2 and 111.6mAhg−1 at current densities of 2C and 5C. After 300 cycles, it still maintained high capacities of 123.5 and 85.1mAhg−1, which corresponds to a capacity loss of 0.07% and 0.08% per cycle, respectively. In addition, even at 10C, it displays a capacity of 68.8mAhg−1 and 87.9% of the capacity (60.5mAhg−1) is remained after 300 cycles. All the results indicate the V2O5 self-assembled nanosheets could be a promising candidate as cathode active material for long-term cycling performance in Lithium-ion batteries.