Journal
NATURE MEDICINE
Volume 24, Issue 10, Pages 1519-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41591-018-0209-1
Keywords
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Funding
- SNSF [31003A_160230]
- Swiss National Science Foundation
- Swiss National Science Foundation (SNF) [31003A_160230] Funding Source: Swiss National Science Foundation (SNF)
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CRISPR-Cas-based genome editing holds great promise for targeting genetic disorders, including inborn errors of hepatocyte metabolism. Precise correction of disease-causing mutations in adult tissues in vivo, however, is challenging. It requires repair of Cas9-induced double-stranded DNA (dsDNA) breaks by homology-directed mechanisms, which are highly inefficient in nondividing cells. Here we corrected the disease phenotype of adult phenylalanine hydroxylase (pah)(enu2) mice, a model for the human autosomal recessive liver disease phenylketonuria (PKU)(1), using recently developed CRISPR-Cas-associated base editors(2-4). These systems enable conversion of C.G to T.A base pairs and vice versa, independent of dsDNA break formation and homology-directed repair (HDR). We engineered and validated an intein-split base editor, which allows splitting of the fusion protein into two parts, thereby circumventing the limited cargo capacity of adeno-associated virus (AAV) vectors. Intravenous injection of AAV-base editor systems resulted in Pah(enu2) gene correction rates that restored physiological blood phenylalanine (L-Phe) levels below 120 mu mol/l [5]. We observed mRNA correction rates up to 63%, restoration of phenylalanine hydroxylase (PAH) enzyme activity, and reversion of the light fur phenotype in Pah(enu2) mice. Our findings suggest that targeting genetic diseases in vivo using AAV-mediated delivery of base-editing agents is feasible, demonstrating potential for therapeutic application.
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