Thermo-responsive block copolymers are of great interest because of their mature synthesis technology, well-defined structure, controllable molecular weight and easy implementation of their nano-assemblies in water by temperature regulation. Up to now, several previous reviews have summarized their application in biomedical fields. Thermo-responsive block copolymers also exhibit promising potential in temperature-regulated adsorption and/or separation, nanoreactors for green organic synthesis, etc. Herein, recent progress in their thermo-responsive assembly in water and application of five kinds of thermo-responsive block copolymers containing one or two thermo-responsive blocks beyond biomedical application is reviewed.