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Fig. 2
gRNA generated containing HH and HDV ribozyme is efficient at inducing mutagenesis of its target site.
(A) mCherryHH-gRNA-HDV construct used to generate gRNA. H2a mCherry is used to label cells expressing gRNA and MALAT is used to stabilize the RNA after cleavage by HH and HDV ribozyme. (B) Injection of Smo gRNA together with Cas9 mRNA causes mutation of its target sequence. (C) Images of smyhc1: GFP embryos at 2dpf show that the majority of GFP expressing cells are lost upon injection of GFP gRNA and Cas9 mRNA. On average, there are ~24 GFP positive slow muscle fibers per somite in uninjected smyhc1: GFP embryos whereas only 6 GFP positive slow muscle fibres remained in each somite of injected smyhc1: GFP embryos (n = 5). Scale bars: 40 μm.