Fig. 2

CRISPR/Cas9-based double gene knockout of PxAPN1 and PxAPN3a in P. xylostella. a The genomic structure of PxAPN1 and PxAPN3a, the sgRNA sequences are highlighted in blue text and the PAM sequences by underlined red text. The disrupted sequences are shown in the gray-blue dashed box. b Genotype detection of double gene mutation in PxAPN1 and PxAPN3a locus according to the banding profile of PCR products from the four primer pairs. c Representative chromatograms of direct sequencing of PCR products with the primer pair 3a-1-F/1-13-R. The arrow indicates the junction of PxAPN1 and PxAPN3a genes after deletion. M: DNA marker III