The Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) is an insect defoliator of the potato, Solanum tuberosum L. This species thrives in agricultural environments because of its flexible and complex life history, as well as its ability to evolve insecticide resistance. As a result, it has become a widely distributed agricultural pest. Ledprona (trade name Calantha) is a recently developed novel double-stranded RNA (dsRNA) insecticide that controls populations of Colorado potato beetle through RNA interference (RNAi). Previous studies have demonstrated the efficacy of ledprona through laboratory, greenhouse, and field studies. Colorado potato beetles from geographically distinct populations are known to vary in their response to insecticides, including experimental compounds based on RNAi. We tested the mortality and foliage consumption of beetles from different areas in the US treated with ledprona and found significant variation in both parameters. The beetles originating from New York were significantly less susceptible to ledprona in leaf disc assays compared to other populations. However, currently there is no evidence of reduced performance of ledprona against that population under field conditions, possibly because intoxicated beetles cannot withstand multiple stressors present in the field. The results of this study confirmed that the ledprona efficacy differs among geographically distinct populations, which may have implications for managing Colorado potato beetles.