In this paper, the absorption and fluorescence spectra of Er3+, Pr3+ co-doped LiYF4 (Er,Pr:YLF) crystal were measured and analyzed. The Pr3+ co-doping was proved to effectively enhance the Er3+:I411/2→I413/2 mid-infrared transition at the 2.7 µm with 74.1% energy transfer efficiency from Er3+:I413/2 to Pr3+:F34. By using the Judd–Ofelt theory, the stimulated emission cross section was calculated to be 1.834×10−20 cm2 at 2685 nm and 1.359×10−20 cm2 at 2804.6 nm. Moreover, a diode-end-pumped Er,Pr:YLF laser operating at 2659 nm was realized for the first time, to the best of our knowledge. The maximum output power was determined to be 258 mW with a slope efficiency of 7.4%, and the corresponding beam quality factors Mx2=1.29 and My2=1.25. Our results suggest that Er,Pr:YLF should be a promising material for 2.7 µm laser generation.