Fig. 2.

Fig. 2.

Killifish and Zebrafish AIP mutants generated by CRISPR-Cas9. A) Top: The gene and mRNA structures of AIP are illustrated. Bottom: All alleles encode frameshift mutations that cause the addition of missense amino acids (red) and premature termination codons, leading to predicted truncated proteins. B) The zebrafish mutant cDNAs were cloned and expressed by in vitro transcription and translation in the presence of [³⁵S]methionine, confirming that they encode truncated proteins. The double band in the middle lane may reflect a partial read-through of the premature termination codon introduced by the frameshift mutation; however, any additional amino acids would be mis-sense residues. Abbreviations: SgRNA: Single guide RNA; FKBP-PPI: FK506-binding protein—peptidyl-prolyl isomerase domain; TPR: Tetratricopeptide; AHR: Aryl hydrocarbon receptor.