Eggs of wild-type AB zebrafish (one-cell stage) were microinjected with 4 ng of the KARS- specific antisense morpholino (MO3i3, TCCATATTCGCTACTCATCGTACA) or with a control morpholino (MOctl, GAAAGCATGGCATCTGGATCATCGA). The KARS-specific MO targets the splice donor site between exon 3 and intron 4. Efficiency of knockdown was assessed by RT-qPCR with splice-sensitive primers [TGGACCCCAATCAATACTTCAAG (forward) and GGTCTCCAGGCTGAAGGTGGTTAT (reverse)]. Embryos then developed with no obvious morphological defects at 28°C. At 72 hours after injection, embryos were collected for gene expression analysis by RT-qPCR.

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