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ATP-binding cassette transporter C2 (ABCC2) is known to be a receptor for Bacillus thuringiensis (Bt) toxins in several lepidopteran insects.Mutations in the ABCC2 gene have been genetically linked to field-evolved resistance to the Cry1F toxin from Bt in Spodoptera frugiperda.Here we generated a SfABCC2 knockout strain of S.frugiperda using the CRISPR/Cas9 system to provide further functional evidence of the role of this gene in susceptibility and resistance to Cry1F.Results from bioassays showed that the SfABCC2 knockout S.frugiperda strain displayed 118-fold resistance to Cry1F compared with the parental DH19 strain,but no resistance to Vip3A toxin from Bt.These results provide the first reverse genetic evidence for SfABCC2 as a functional receptor for Cry1F.