Hydrostatic pressure is an important environmental factor affecting the vertical distribution of marine organisms. Laboratory-based studies have shown that many extant shallow-water marine benthic invertebrates can tolerate hydrostatic pressure outside their known natural distributions. However, only a few studies have focused on the molecular mechanisms of pressure acclimatisation. In the present work, we examined the pressure tolerance of the shallow-water amphipod Eogammarus possjeticus at various temperatures (5, 10, 15, and 20 °C) and hydrostatic pressures (0.1-30 MPa) for 16 h. Six of these experimental groups were used for transcriptome analysis. We found that 100% of E. possjeticus survived under 20 MPa at all temperature conditions for 16 h. Sequence assembly resulted in 138, 304 unigenes. Results of differential expression analysis revealed that 94 well-annotated genes were up-regulated under high pressure. All these findings indicated that the pressure tolerance of E. possjeticus was related to temperature. Several biological processes including energy metabolism, antioxidation, immunity, lipid metabolism, membrane-related process, genetic information processing, and DNA repair are probably involved in the acclimatisation in deep-sea environments.