1. Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.
- Author
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Smith LJ, Wright J, Clark G, Ul-Hasan T, Jin X, Fong A, Chandra M, St Martin T, Rubin H, Knowlton D, Ellsworth JL, Fong Y, Wong KK Jr, and Chatterjee S
- Subjects
- BRCA2 Protein physiology, Genetic Vectors, Humans, Interleukin Receptor Common gamma Subunit genetics, K562 Cells, Dependovirus genetics, Gene Editing, Hematopoietic Stem Cells metabolism, Homologous Recombination
- Abstract
The precise correction of genetic mutations at the nucleotide level is an attractive permanent therapeutic strategy for human disease. However, despite significant progress, challenges to efficient and accurate genome editing persist. Here, we report a genome editing platform based upon a class of hematopoietic stem cell (HSC)-derived clade F adeno-associated virus (AAV), which does not require prior nuclease-mediated DNA breaks and functions exclusively through BRCA2-dependent homologous recombination. Genome editing is guided by complementary homology arms and is highly accurate and seamless, with no evidence of on-target mutations, including insertion/deletions or inclusion of AAV inverted terminal repeats. Efficient genome editing was demonstrated at different loci within the human genome, including a safe harbor locus, AAVS1, and the therapeutically relevant IL2RG gene, and at the murine Rosa26 locus. HSC-derived AAV vector (AAVHSC)-mediated genome editing was robust in primary human cells, including CD34
+ cells, adult liver, hepatic endothelial cells, and myocytes. Importantly, high-efficiency gene editing was achieved in vivo upon a single i.v. injection of AAVHSC editing vectors in mice. Thus, clade F AAV-mediated genome editing represents a promising, highly efficient, precise, single-component approach that enables the development of therapeutic in vivo genome editing for the treatment of a multitude of human gene-based diseases., Competing Interests: Conflict of interest statement: S.C. and L.J.S. are cofounders of Homology Medicines, Inc., and S.C., L.J.S., J.W., T.S.M., H.R., J.L.E., and D.K. hold equity in Homology Medicines, Inc.- Published
- 2018
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