CRISPR-Cas9 induced lrp5 zebrafish knockout line. (A) Schematic representation of the ZF lrp5 gene structure with Sanger sequence chromatogram; (B) demonstrating the del8bp (Ala53fs) mutation, that results in a frameshift and a premature stops codon; (C) Schematic representation of the ZF LRP5 wild-type and mutated protein demonstrates the lack of functional domains in LRP5 of lrp5-/- (LY -low-density lipoprotein receptor; EGF – epidermal growth factor; LDLa- Low-density lipoprotein receptor domain class (A, D) Western-blot results demonstrated the absence of LRP5 protein in lrp5-/-.(E) Each column in the graph represents the proportion of genotype (%) per developmental stage. Only 3.7% of lrp5-/- in survive and reach adolescence using instead of expected 25%.
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