PAH deficient pathology in humanized c.1066-11G>A phenylketonuria mice

Hum Mol Genet. 2024 Jun 5;33(12):1074-1089. doi: 10.1093/hmg/ddae051.


We have generated using CRISPR/Cas9 technology a partially humanized mouse model of the neurometabolic disease phenylketonuria (PKU), carrying the highly prevalent PAH variant c.1066-11G>A. This variant creates an alternative 3' splice site, leading to the inclusion of 9 nucleotides coding for 3 extra amino acids between Q355 and Y356 of the protein. Homozygous Pah c.1066-11A mice, with a partially humanized intron 10 sequence with the variant, accurately recapitulate the splicing defect and present almost undetectable hepatic PAH activity. They exhibit fur hypopigmentation, lower brain and body weight and reduced survival. Blood and brain phenylalanine levels are elevated, along with decreased tyrosine, tryptophan and monoamine neurotransmitter levels. They present behavioral deficits, mainly hypoactivity and diminished social interaction, locomotor deficiencies and an abnormal hind-limb clasping reflex. Changes in the morphology of glial cells, increased GFAP and Iba1 staining signals and decreased myelinization are observed. Hepatic tissue exhibits nearly absent PAH protein, reduced levels of chaperones DNAJC12 and HSP70 and increased autophagy markers LAMP1 and LC3BII, suggesting possible coaggregation of mutant PAH with chaperones and subsequent autophagy processing. This PKU mouse model with a prevalent human variant represents a useful tool for pathophysiology research and for novel therapies development.

Keywords: CRISPR/Cas; autophagy; humanized mouse model; phenylketonuria; splice variant.

MeSH terms

  • Animals
  • Autophagy / genetics
  • Brain / metabolism
  • Brain / pathology
  • CRISPR-Cas Systems
  • Disease Models, Animal*
  • Humans
  • Liver / metabolism
  • Liver / pathology
  • Mice
  • Mutation
  • Phenylalanine Hydroxylase* / genetics
  • Phenylalanine Hydroxylase* / metabolism
  • Phenylketonurias* / genetics
  • Phenylketonurias* / metabolism
  • Phenylketonurias* / pathology


  • Phenylalanine Hydroxylase