A porcine model of phenylketonuria generated by CRISPR/Cas9 genome editing
| Autor: | Robert Wagner, Kevin D. Wells, Steven F. Dobrowolski, Susanne M. Gollin, Robert D. Nicholls, Yijen L. Wu, Marie A. Johnson, Eric M. Walters, Erik A. Koppes, Sarah A. Hansen, M Yerle, Shawn E. Christ, Dale W. Lewis, Melissa Samuel, Joseph T. Newsome, Kristen J. Skvorak, Megan E. Yates, Lina Ghaloul-Gonzalez, Randall S. Prather, Uta Lichter-Konecki, Stephanie L. Murphy, Lee D. Spate, Joshua A. Benne, Jerry Vockley, Bethany K. Redel, Angela Leshinski |
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| Přispěvatelé: | Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Adult
0301 basic medicine Adolescent Swine Phenylalanine [SDV]Life Sciences [q-bio] Biology Amino acid metabolism Mouse models Compound heterozygosity 03 medical and health sciences Exon 0302 clinical medicine Hyperphenylalaninemia Phenylketonurias Genetics medicine Animals Humans CRISPR Gene Editing Zygote Neurotoxicity Phenylalanine Hydroxylase General Medicine medicine.disease Phenotype Diet 3. Good health Disease Models Animal Metabolism 030104 developmental biology Liver 030220 oncology & carcinogenesis Medicine CRISPR-Cas Systems Research Article Genetic diseases |
| Zdroj: | JCI Insight JCI Insight, American Society for Clinical Investigation, 2020, 5 (20), ⟨10.1172/jci.insight.141523⟩ JCI Insight, Vol 5, Iss 20 (2020) |
| ISSN: | 2379-3708 |
| DOI: | 10.1172/jci.insight.141523⟩ |
| Popis: | Phenylalanine hydroxylase–deficient (PAH-deficient) phenylketonuria (PKU) results in systemic hyperphenylalaninemia, leading to neurotoxicity with severe developmental disabilities. Dietary phenylalanine (Phe) restriction prevents the most deleterious effects of hyperphenylalaninemia, but adherence to diet is poor in adult and adolescent patients, resulting in characteristic neurobehavioral phenotypes. Thus, an urgent need exists for new treatments. Additionally, rodent models of PKU do not adequately reflect neurocognitive phenotypes, and thus there is a need for improved animal models. To this end, we have developed PAH-null pigs. After selection of optimal CRISPR/Cas9 genome-editing reagents by using an in vitro cell model, zygote injection of 2 sgRNAs and Cas9 mRNA demonstrated deletions in preimplantation embryos, with embryo transfer to a surrogate leading to 2 founder animals. One pig was heterozygous for a PAH exon 6 deletion allele, while the other was compound heterozygous for deletions of exon 6 and of exons 6–7. The affected pig exhibited hyperphenylalaninemia (2000–5000 μM) that was treatable by dietary Phe restriction, consistent with classical PKU, along with juvenile growth retardation, hypopigmentation, ventriculomegaly, and decreased brain gray matter volume. In conclusion, we have established a large-animal preclinical model of PKU to investigate pathophysiology and to assess new therapeutic interventions. A phenylalanine hydroxylase–deficient pig model of phenylketonuria is established and characterized. |
| Databáze: | OpenAIRE |
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