Development of a porcine model of phenylketonuria with a humanized R408W mutation for gene editing

Phenylketonuria (PKU) is a metabolic disorder whereby phenylalanine metabolism deficient due to allelic variations in the gene for hydroxylase ( PAH ). There no cure PKU other than orthotopic liver transplantation, and standard of care patients limited dietary restrictions key amino acid supplementation. Therefore, Pah was edited pig fibroblasts generation clone piglets that harbor common severe human mutation, R408W. Additionally, proximal region mutation further humanized by introducing 5 single nucleotide polymorphisms (SNPs) allow development editing machinery could be translated directly from model at least one classic R408W allele. Resulting were hypopigmented (a Ossabaw piglet) had low birthweight (all piglets). The similar levels expression, but detectable enzymatic activity, consistent with phenotype. fragile required extensive neonatal prevent failure thrive early demise. Phenylalanine rose sharply when Phe unrestricted rapidly reduced diet. Fibroblasts isolated show susceptibility correction using CRISPR or TALEN, subsequent homology-directed recombination correct . This provides powerful new tool all classes therapeutic candidates treat PKU, as well unique value proof-of-concept studies vivo platforms context this