Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing
| Autor: | Laura J. Smith, Jason Wright, Gabriella Clark, Taihra Ul-Hasan, Xiangyang Jin, Abigail Fong, Manasa Chandra, Thia St Martin, Hillard Rubin, David Knowlton, Jeff L. Ellsworth, Yuman Fong, Kamehameha K. Wong, Saswati Chatterjee |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Genetic Vectors Locus (genetics) Computational biology Biology medicine.disease_cause Corrections Homology (biology) 03 medical and health sciences Genome editing medicine Humans Homologous Recombination Gene Adeno-associated virus BRCA2 Protein Gene Editing Multidisciplinary Hematopoietic stem cell Dependovirus Hematopoietic Stem Cells 030104 developmental biology medicine.anatomical_structure Human genome Homologous recombination K562 Cells Interleukin Receptor Common gamma Subunit |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 115(31) |
| ISSN: | 1091-6490 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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