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The classic  brown body  ( bwb ) mutation in the housefly  Musca domestica  impairs normal melanization of the adult cuticle. In  Drosophila melanogaster , a reminiscent pigmentation defect results from mutations in the  yellow  gene encoding dopachrome conversion enzyme (DCE). Here, we demonstrate that the  bwb  locus structurally and functionally represents the  yellow  ortholog of  Musca domestica ,  MdY . In  bwb Musca  strains, we identified two mutant  MdY  alleles that contain lesions predicted to result in premature truncation of the MdY open reading frame. We targeted wildtype  MdY  by CRISPR-Cas9 RNPs and generated new mutant alleles that fail to complement existing  MdY  alleles, genetically confirming that  MdY  is the  bwb  locus. We further found evidence for Cas9-mediated interchromosomal recombination between wildtype and mutant  bwb  alleles. Our work resolves the molecular identity of the classic  bwb  mutation in  Musca domestica  and establishes the feasibility of Cas9-mediated genome editing in the  Musca  model.

CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus