ZFIN Image: Kolvenbach et al., 2019, Fig. S4

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Figure Caption

Fig. S4

Transient Bnc2 CRISPR/Cas9 Knockdown in Zebrafish Larvae Reproduces the Morpholino Phenotype

(A): Schematic showing the genomic positions of the different guide-RNAs used for the transient Bnc2 CRISPR/Cas9 targeting experiments.

(B-C): Single guide RNA (sgRNA) injections of guide 6 targeting bnc2 with Cas9 reproduces the distal ‘vesicle’ as an outlet obstruction of the pronephric ducts in 5 % of injected embryos at 33 hpf (see inlays - black arrowheads). Kinked tails were observed in 1 % of the zfl (black asterisk).

(D): Follow-up of sgRNA injected zfl targeting bnc2 for sgRNA 4 shows hydrocephalus (black asterisk) and light body curvature (black arrowheads) at 54 hpf.

(E): Kaplan-Meier plots for survival of acute multi sgRNA (mgRNA) approach for guides 2, 6 and 8 targeting bnc2 with Cas9. Zfl were included in survival experiment after 24 hpf. Injected larvae show significantly (p<0.0001, Mantel-Cox test) reduced survival (median survival of 7 dpf) and 40 % survival at the end of the experiment.

Scale bars: 500 μm (B, C and D)

Acknowledgments

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