We demonstrate single-mode laser operation in dye-doped holographic polymer-dispersed liquid crystal (HPDLC) transmission gratings. The gratings are fabricated in cells made from specifically chosen glass substrates to decrease the refractive index difference between the waveguide core layer and cladding layer. The phase separation degree of liquid crystal after holographic recording is further optimized to confine only the lowest propagation mode in the device. The mode selection mechanism is explained under the framework of the waveguide distributed feedback (DFB) theory. The wavelength of single-mode lasing can be tuned between 620 and 660 nm by varying the grating period. Our results show the HPDLC technique could provide single-mode organic DFB lasers in a tunable, simple, and large-area manner.