Morpholino oligonucleotides (MOs) used for microinjection were ordered from Gene Tools. The MOs were microinjected into one-cell-stage embryos as previously described (Nasevicius and Ekker, 2000 (link)). Transient tprb transgene construct within Tol2 vectors (40 pg) was microinjected into one-cell-stage embryos with Tol2 transposase mRNA (50 pg). CRISPR-Cas9–mediated generation of zebrafish mutants (tprb–3+1 and vhl) were performed as previously described (Xiao et al., 2013 (link)). The gRNAs (50 pg) and Cas9 protein (500 pg) were co-microinjected into one-cell-stage embryos.
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