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Construction of the CRISPR/Cas9 system of transgenic silkworm. Transgenic vector construction of pBac[IE1-Cas9-Ser-PA-3 × P3 EGFP afm] and pBac[U6-sgRNA-3 × P3 DsRed afm] injection with the helper plasmid. G1 generation of Cas9- and sgRNA-positive strains were detected by fluorescence screening, G2 generation four transgenic hybrid lines, namely, Cas9(–)/sgRNA(–), Cas9(+)/sgRNA(–), Cas9(+)/sgRNA(–), and Cas9(+)/sgRNA(+) were generated by G1 hybridization.
