Fig. 2

(A) Dazl protein diagram illustrating the RRM and DAZ domains with their respective amino acid regions. ZFN and CRISPR sites are indicated by red lines. (B) dazl alleles generated by ZFN (dazl^Δ7) and CRISPR (dazl^ae57and dazl^ae34). Partial ZFNs and Cas9 binding sites are highlighted in red. Dashed lines represent deletions and orange denotes substitutions. (C) Diagram of alleles generated. The ZFN allele (top) causes a 7 bp deletion and leads to a premature stop codon eliminating all functional domains. dazl^ae57 (second from top) creates a 9 bp insertion and 2 bp substitution leading to a premature stop codon after the RRM domain. The third allele, induced a 12 bp deletion and 15 bp insertion causing an in-frame deletion between the RRM and DAZ domains. (D) Representative PCR/derived cleaved amplified polymorphic sequences genotyping assay for each allele. Heterozygotes harbor the wild-type allele (upper band) and the mutant allele (lower band) in the case of dazl^Δ7. dazl^ae57 or dazl^ae34 heterozygotes harbor wild-type (lower band) and mutant alleles (upper band).