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Phenotypes in the mrps2 knockout zebrafish larvae. A A schematic of the zebrafish mrps2 with the location of the selected sgRNA target sites indicated. The sequences of the two sgRNAs used in the study are provided in the table, B Assessment of editing efficiency for each sgRNA at 24 h post injection, by a heteroduplex mobility assay (HMA), C Relative expression levels of the mrps2 mRNA in the controls and injectants (crispants), D Representative bright field images of wild-type and mrps2 crispants (at 3dpf) illustrating abnormal developmental phenotypes observed in the crispants, as labeled, E, F Quantification of the number of larvae showing abnormal development at 5dpf (E) and survival at 7dpf (F) (5 experiments, NT n = 560, KO n = 820), G Complex IV activity measurement in mitochondria isolated from control and mrps2 crispants, H 12S to 16S rRNA ratio in the crispants compared to controls. Results from 3 to 5 independent experiments are quantified, and error bars represent SEM, (I) Relative mRNA levels of OXPHOS subunits genes including nd1, ndufs1 (Complex I), sdha (Complex II), cytb (Complex III), mt-co1 and mt-co2 (Complex IV), in the mrps2 crispants compared to controls.
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