CRISPR-Cas9 Targeting of Sf-FGFR and Sf-SR-C

The sgRNA of Sf-SR-C, Sf-FGFR gene was designed using sgRNA design software, and the off-target risk of sgRNA was evaluated using the whole genome of S. frugiperda as a reference sequence [34 (link)]. The N18NGG sequence with the highest score and no potential off-target sites was selected as the sgRNA target sequence. The Sf-FGFR target sequence (5′-TATGTGCAGCAGAAACATTGG-3′, the PAM sequence is underlined) at exon 1 of Sf-FGFR, and the Sf-SR-C target sequence (5′-TGTACTGTGATGGATCCAATTGG-3′, the PAM sequence is underlined) at exon 1 of Sf-SR-C. SgRNA was synthesized in vitro according to the instructions of the GeneArt™ Precision gRNA Synthesis Kit. Then the sgRNA was purified by the gRNA Clean Up Kit. Cas9 protein (GeneArt ™ Platinum ™ Cas9 Nuclease) was purchased from Thermo Fisher Scientific (Shanghai, China).