Total RNA from leaf tissue of glasshouse-cultivated plants was extracted as described previously (60). Total nucleic acid (2 μg), determined using NanoDrop, was treated with deoxyribonuclease, and a complementary DNA (cDNA) library was prepared using the RevertAid First Strand cDNA Synthesis Kit following the manufacturer’s guidelines (Thermo Fisher Scientific). Oligonucleotide primers specific to AtFT (qPCR-FT-F and qPCR-FT-R), GBSSsgRNA4 (GBSSsg4-F and sgRNA-R), PTSTsgRNA2 (PTSTsg2-F and sgRNA-R), and Cas9 (Cas9-F1 and Cas9-R) were used with 1.5 μl of cDNA and Fast SYBR Green Master Mix according to the manufacturer’s guidelines (Applied Biosystems). Data were analyzed using the 2−ΔCt equation (61), and transgene expression was determined relative to the endogenous MePP2A reference gene amplified using oligonucleotide primers PP2A-LP2 and PP2A-RP2.

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