Figure 1.

Cyp4v3 gene structure, mutations, and expression in the KO mouse model. (A). Schematic of the mouse Cyp4v3 gene showing primers used for amplification and sequence tracing, as shown for the two insertions induced in the Cyp4v3 gene. (B). Novel transcript and associated gene structure in KO mice, including an extra 137 bp exon (labeled 2b) between exons 2 and 3 and skipped exon 4. (C). RT-PCR of Cyp4v3 mRNA spanning exons 2 to 9. The red arrow indicates the novel transcript band, including exon 2b, with reverse sequencing results shown in (D), with dual sequences of exons 2 and 2b in the region marked exon 2. The black arrow indicates the band for the transcript skipping exon 4, which is present in both the KO and, to a lesser extent, the wild-type sequences, the sequence trace of which is shown in (E,F). The exact sequence for the novel transcript in the KO mice with the exon 2b sequence (shaded blue) and TGA codons (underlined in red), showing the premature stop codons in exons 2b and 3.