Treatment of a metabolic liver disease in mice with a transient prime editing approach

Prime editing is a versatile genome editing technology that circumvents the need for DNA double-strand break formation and homology-directed repair, making it particularly suitable for in vivo correction of pathogenic mutations. Here we developed liver-specific prime editing approaches with temporally restricted prime editor (PE) expression. We first established a dual-delivery approach where the prime editor guide RNA is continuously expressed from adeno-associated viral vectors and only the PE is transiently delivered as nucleoside-modified mRNA encapsulated in lipid nanoparticles (LNP). This strategy achieved 26.2% editing with PEmax and 47.4% editing with PE7 at the Dnmt1 locus using a single 2 mg kg<sup>-1</sup> dose of mRNA-LNP. When targeting the pathogenic Pah<sup>enu2</sup> mutation in a phenylketonuria mouse model, gene correction rates reached 4.3% with PEmax and 20.7% with PE7 after three doses of 2 mg kg<sup>-1</sup> mRNA-LNP, effectively reducing blood L-phenylalanine levels from over 1,500 µmol l<sup>-1</sup> to below the therapeutic threshold of 360 µmol l<sup>-1</sup>. Encouraged by the high efficiency of PE7, we next explored a simplified approach where PE7 mRNA was co-delivered with synthetic prime editor guide RNAs encapsulated in LNP. This strategy yielded 35.9% editing after two doses of RNA-LNP at the Dnmt1 locus and 8.0% editing after three doses of RNA-LNP at the Pah<sup>enu2</sup> locus, again reducing L-phenylalanine levels below 360 µmol l<sup>-1</sup>. These findings highlight the therapeutic potential of mRNA-LNP-based prime editing for treating phenylketonuria and other genetic liver diseases, offering a scalable and efficient platform for future clinical translation.

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Publication Details

Journal
Nature Biomedical Engineering
Published
2025-05-20
DOI
https://doi.org/10.1038/s41551-025-01399-4
Citations
12
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
9.67

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article

Treatment of a metabolic liver disease in mice with a transient prime editing approach

Lukas Villiger, Gerald Schwank, Kim Fabiano Marquart, Lukas Schmidheini et al.
12 citations
Nature Biomedical Engineering
CRISPR and Genetic Engineering
9.67
article

Treatment of a metabolic liver disease in mice with a transient prime editing approach

Lukas Villiger, Gerald Schwank, Kim Fabiano Marquart, Lukas Schmidheini, Paulo J.C. Lin, Beat Thöny, Péter István Kulcsár, Steven H.Y. Fan, Tanja Rothgangl, Johannes Häberle, Mai Matsushita, Nicole Rimann, Ying K. Tam, Norbert Pardi, Alessio Cremonesi, Yanik Weber, Hiromi Muramatsu, András Tálas, Máté Vadovics, Erica Faccin, Manfred Köpf, Elina Andrea Villiger, Eleonora I. Ioannidi, Desirée Böck, Cornelia Schwerdel, Woohyun J Moon
article en
12 citations

Abstract

Prime editing is a versatile genome editing technology that circumvents the need for DNA double-strand break formation and homology-directed repair, making it particularly suitable for in vivo correction of pathogenic mutations. Here we developed liver-specific prime editing approaches with temporally restricted prime editor (PE) expression. We first established a dual-delivery approach where the prime editor guide RNA is continuously expressed from adeno-associated viral vectors and only the PE is transiently delivered as nucleoside-modified mRNA encapsulated in lipid nanoparticles (LNP). This strategy achieved 26.2% editing with PEmax and 47.4% editing with PE7 at the Dnmt1 locus using a single 2 mg kg<sup>-1</sup> dose of mRNA-LNP. When targeting the pathogenic Pah<sup>enu2</sup> mutation in a phenylketonuria mouse model, gene correction rates reached 4.3% with PEmax and 20.7% with PE7 after three doses of 2 mg kg<sup>-1</sup> mRNA-LNP, effectively reducing blood L-phenylalanine levels from over 1,500 µmol l<sup>-1</sup> to below the therapeutic threshold of 360 µmol l<sup>-1</sup>. Encouraged by the high efficiency of PE7, we next explored a simplified approach where PE7 mRNA was co-delivered with synthetic prime editor guide RNAs encapsulated in LNP. This strategy yielded 35.9% editing after two doses of RNA-LNP at the Dnmt1 locus and 8.0% editing after three doses of RNA-LNP at the Pah<sup>enu2</sup> locus, again reducing L-phenylalanine levels below 360 µmol l<sup>-1</sup>. These findings highlight the therapeutic potential of mRNA-LNP-based prime editing for treating phenylketonuria and other genetic liver diseases, offering a scalable and efficient platform for future clinical translation.

Nature Biomedical EngineeringVol. 9(10)
University of Zurich (CH), ETH Zurich (CH), University Children's Hospital Zurich (CH), Acuitas Therapeutics (Canada) (CA), University of Pennsylvania (US)
National Science Foundation, Novartis Foundation, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, Eidgenössische Technische Hochschule Zürich, Novartis Stiftung für Medizinisch-Biologische Forschung, Universität Zürich, National Institutes of Health, Staatssekretariat für Bildung, Forschung und Innovation, National Institute of Allergy and Infectious Diseases
Good health and well-being
Openalex Percentile: Top 2%
CRISPR and Genetic Engineering
9.67
DOIOpen Access PDFOpenAlex
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