Fig. 2

CRISPR-induced indels can be detected in the late zebrafish blastula. a RNAs encoding the Cas9 endonuclease and a guide RNA (gRNA) targeting gldc coding sequence are co-microinjected into one-cell stage embryos. The genomic DNA was extracted either at the sphere stage (4 h post-fertilization, hpf) or at the prim-6 stage (24 hpf) using the raw extraction method. b Sequencing electropherograms of 8 wild type and 8 CRISPR/Cas9-injected embryos at 24 hpf. The black arrow shows the presence of faint multiple peaks in the CRISPR/cas9-injected embryos, indicating the presence of indels. c HRM curve analysis of uninjected wild type (WT, red curves) and CRISPR/Cas9-injected (green curves) embryos at 4 hpf. The green curves are shifted and irregular compared to the red WT curves, indicating the presence of indels at the late blastula stage. d Melting curve analysis of uninjected WT (red curves) and CRISPR/Cas9-injected (green curves) embryos at 24 hpf