In vivo expansion of gene-targeted hepatocytes through transient inhibition of an essential gene
Homology-directed repair (HDR)-based genome editing is an approach that could permanently correct a broad range of genetic diseases. However, its utility is limited by inefficient and imprecise DNA repair mechanisms in terminally differentiated tissues. Here, we tested Repair Drive, a platform technology for selectively expanding HDR-corrected hepatocytes in adult mice in vivo. Repair Drive involves transient conditioning of the liver by knocking down an essential gene, <i>fumarylacetoacetate hydrolase</i> (<i>Fah</i>), and delivering an untargetable version of the essential gene in cis with a therapeutic transgene. We show that Repair Drive increased the percentage of correctly targeted hepatocytes in healthy wild-type mice up to 25%, which resulted in a fivefold increased expression of a therapeutic transgene, <i>human factor IX</i> (<i>FIX</i>). Repair Drive was well tolerated and did not induce toxicity or tumorigenesis during a 1-year follow-up. This approach may broaden the range of liver diseases that can be treated with somatic genome editing.
Authors
- Sarah Hyde
- June Qin
- Lavanya Saxena (ORCID: https://orcid.org/0000-0001-6083-4013)
- Christine Beeton (ORCID: https://orcid.org/0000-0002-2500-5874)
- Karl‐Dimiter Bissig (ORCID: https://orcid.org/0000-0002-9577-9250)
- Tyler Chickering
- M. Cecilia Ljungberg (ORCID: https://orcid.org/0000-0003-0544-878X)
- Kalyani R. Patel (ORCID: https://orcid.org/0000-0003-2654-5386)
- Jun Han (ORCID: https://orcid.org/0000-0002-9477-8782)
- Ayrea Hurley (ORCID: https://orcid.org/0000-0002-8697-8610)
- So Hyun Park (ORCID: https://orcid.org/0000-0003-3599-1687)
- William R. Lagor (ORCID: https://orcid.org/0000-0002-1703-5125)
- Alexandria M. Doerfler (ORCID: https://orcid.org/0000-0003-2297-6103)
- Marcel Chuecos (ORCID: https://orcid.org/0000-0002-1840-0948)
- Kelly Wassarman
- Vasant Jadhav (ORCID: https://orcid.org/0000-0002-6666-0250)
- Adam Castoreno
- Gang Bao (ORCID: https://orcid.org/0000-0001-5501-554X)
- Marco De Giorgi (ORCID: https://orcid.org/0000-0002-9823-1397)
- Mark K. Schlegel (ORCID: https://orcid.org/0000-0002-0127-608X)
- Mia N. Furgurson
- Christopher J. Walkey (ORCID: https://orcid.org/0000-0002-1471-0447)
- Ivan Zlatev (ORCID: https://orcid.org/0000-0003-2797-3032)
- James F. Martin (ORCID: https://orcid.org/0000-0002-7842-9857)
- Mingming Cao (ORCID: https://orcid.org/0000-0002-4234-1175)
- Stéphanie Lefebvre (ORCID: https://orcid.org/0000-0003-4833-2197)
- Jong Kim (ORCID: https://orcid.org/0000-0002-0257-0547)
- Gang Li (ORCID: https://orcid.org/0000-0001-9661-8304)
- Patrick J. Miller (ORCID: https://orcid.org/0000-0002-8418-127X)
Institutions
- Alnylam Pharmaceuticals (United States) (US)
- Duke University (US)
- Baylor College of Medicine (US)
- University of Victoria (CA)
- Center for Human Genetics (US)
- Neurological Research Institute (US)
- Texas Children's Hospital (US)
- Rice University (US)
Publication Details
- Journal
- Science Translational Medicine
- Published
- 2025-02-12
- DOI
- https://doi.org/10.1126/scitranslmed.adk3920
- Citations
- 6
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 2.42