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Fig. 1 Generation of zebrafish Stat5.1 mutants using CRISPR/Cas9. (A) Schematic diagram of STAT5B/Stat5.1 and its constituent N-terminal domain (NTD), coiled-coil domain (CCD), DNA-binding domain (DBD), linker domain (LD), Src-homology 2 domain (SH2), tyrosine residue (Y), and transactivation domain (TAD). An amino acid alignment for the indicated region of Homo sapiens (Hs) STAT5B and Danio rerio (Dr) Stat5.1 is shown above, with identical residues indicated by asterisks. (B) Part of the zebrafish stat5.1 gene. Exons are shown as boxes in color matching the corresponding domain, along with the sequence targeted by the sgRNA. (C) Sequence traces, corresponding nucleotides, and encoded amino acids for homozygous wild-type (WT) (wt/wt), Δ TAD (mdu033/mdu033) and Δ TM (mdu032/mdu032) Stat5.1. The dotted boxes on the WT trace indicate the sequences deleted for the specified mutant. The mdu033 allele represents a 4 bp deletion leading to an altered reading frame after P701 that results in 3 de novo residues followed by a stop codon (shown in red), whereas the mdu032 allele denotes a 12 bp in-frame deletion that results in the removal of 4 residues after K700 with all remaining C-terminal sequences intact. (D) Schematic representation of Stat5.1 WT along with the Δ TAD, Δ TM, and KO (mdu022/mdu022) mutants.