Figure 1 | Scientific Reports

Figure 1

From: CRISPR/Cas9 generated knockout mice lacking phenylalanine hydroxylase protein as a novel preclinical model for human phenylketonuria

Figure 1

Creation of a Pah-KO mouse model. (A) The mouse genomic locus on chromosome 10 and the murine targeted allele after CRISPR gene editing are shown. Exon 1 contains the translation initiation codon. NCBI GeneID: 18478; RefSeq transcript: NM_008777. Exon 1 sequence is shadowed gray, the start codon (ATG) is framed, gRNA is shown as white letters on a black background, the asterisks and arrow indicate the position of PAM and G>T transition. The donor oligo for CRISPR targeting is in bold and underlined. (B) Sequencing genomic DNA samples of the founder 5349. The point mutation of G to T is indicated in the black boxes. (C) Western blot analyses to detect PAH protein in liver or kidney of wildtype (wt) or Hom (ko) mice. Solubilized proteins (60 µg) from mouse liver or kidney were fractionated and immune-blotted with an anti-PAH antibody. The PAH protein (red; expected size 50 kDa) is evident in wildtype mice (lanes 1, 3, 5 and 7) but is absent in Hom mice (2, 4, 6 and 8); lanes 1, 2, 5, 6) are from kidney; lanes 3, 4, 7, 8 are from liver. Equal loading is indicated by probing with antibody to beta-actin (green). M, molecular weight marker lane (bands not visible in this image).

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