<i>lrp5</i> CRISPR/Cas9 injected embryos display normal CNCC induction, but defective CNCC cell migration.

<p>(A) RFLP analysis of 15 pooled embryos per sample. Both, <i>lrp5</i> CRISPR1 and CRISPR2 generate mutations indicated by presence of undigested mutant bands after <i>Not</i>I and <i>Bgl</i>II digestion (right lanes), respectively. Left lanes show non-digested (ND) PCR products. (B-E) In situ hybridization showing <i>foxd3</i> expression at 10 ss in wild-type embryos (B, C) and <i>lrp5</i> CRISPR1 injected embryos (D,E). (F-Q) <i>crestin</i> expression at 20 ss in wild-type embryos (F-I), <i>lrp5</i> CRISPR1 injected embryos (J-M), and <i>lrp5</i> CRISPR2 injected embryos (N-Q). (R-U) Combined bone and cartilage staining at 7 dpf in wild-type (R) and <i>lrp5</i> CRISPR1 and CRISPR2 co-injected embryos showing cartilage defects with different degrees of severity. Note absence of ceratobranchials 1–4 (in S; arrow), and flipped ceratohyal (in T,U; arrows). (B,D,F,J,N) Lateral views with anterior to the left. All remaining images are dorsal views, except (R-U), which are ventral views. Higher magnification views of areas boxed in (G,K,O) are shown in accompanying images (I, M and Q, white boxes) and (H, L and P, black boxes). Note ectopic <i>crestin</i> positive cells in dorsal neuroepithelium (K,L,O,P) and reduced migratory CNCCs streams (K,M,O,Q) in <i>lrp5</i> CRISPR injected embryos. Scale bars: 100 μm (D,G,N) and 50μm (E,H,I).