Induced Liver Regeneration Enhances CRISPR/Cas9-Mediated Gene Repair in Tyrosinemia Type 1
| Autor: | Qing Shuo Zhang, Sean Nygaard, Niveditha Balaji, Kevin Baradar, Markus Grompe, Amita Tiyaboonchai, Angela Major |
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| Rok vydání: | 2020 |
| Předmět: |
Genetic enhancement
Biology Tyrosinemia 03 medical and health sciences Mice 0302 clinical medicine Fanconi anemia Genetics medicine CRISPR Animals Molecular Biology Research Articles 030304 developmental biology Gene Editing 0303 health sciences Cas9 Tyrosinemias Genetic Therapy medicine.disease Liver regeneration Liver Regeneration 030220 oncology & carcinogenesis Mutation (genetic algorithm) Cancer research Hepatocytes Molecular Medicine CRISPR-Cas Systems Homologous recombination |
| Zdroj: | Hum Gene Ther |
| ISSN: | 1557-7422 |
| Popis: | The efficiency of gene repair by homologous recombination in the liver is enhanced by CRISP/Cas9 incision near the mutation. In this study, we explored interventions designed to further enhance in vivo hepatocyte gene repair in a model of hereditary tyrosinemia. A two-AAV system was employed: one virus carried a Staphylococcus pyogenes Cas9 (SpCas9) expression cassette and the other harbored a U6 promoter-driven sgRNA and a fragment of fumarylacetoacetate hydrolase (Fah) genomic DNA as the homologous recombination donor. In neonatal mice, a gene correction frequency of ∼10.8% of hepatocytes was achieved. The efficiency in adult mice was significantly lower at ∼1.6%. To determine whether hepatocyte replication could enhance the targeting frequency, cell division was induced with thyroid hormone T3. This more than doubled the gene correction efficiency to 3.5% (p |
| Databáze: | OpenAIRE |
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