Journal article
High-efficiency transgene integration by homology-directed repair in human primary cells using DNA-PKcs inhibition
Nature biotechnology, Vol.42(5), pp.731-744
05/01/2024
DOI: 10.1038/s41587-023-01888-4
PMID: 37537500
Abstract
Therapeutic applications of nuclease-based genome editing would benefit from improved methods for transgene integration via homology-directed repair (HDR). To improve HDR efficiency, we screened six small-molecule inhibitors of DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a key protein in the alternative repair pathway of non-homologous end joining (NHEJ), which generates genomic insertions/deletions (INDELs). From this screen, we identified AZD7648 as the most potent compound. The use of AZD7648 significantly increased HDR (up to 50-fold) and concomitantly decreased INDELs across different genomic loci in various therapeutically relevant primary human cell types. In all cases, the ratio of HDR to INDELs markedly increased, and, in certain situations, INDEL-free high-frequency (>50%) targeted integration was achieved. This approach has the potential to improve the therapeutic efficacy of cell-based therapies and broaden the use of targeted integration as a research tool.
A small molecule enhances targeted gene integration at therapeutically relevant loci in human primary cells.
Details
- Title: Subtitle
- High-efficiency transgene integration by homology-directed repair in human primary cells using DNA-PKcs inhibition
- Creators
- Sridhar Selvaraj - Stanford UniversityWilliam N. Feist - Stanford UniversitySebastien Viel - Stanford UniversitySriram Vaidyanathan - Stanford UniversityAmanda M. Dudek - Stanford UniversityMarc Gastou - Stanford UniversitySarah J. Rockwood - Stanford UniversityFreja K. Ekman - Stanford UniversityAluya R. Oseghale - Stanford UniversityLiwen Xu - Stanford UniversityMara Pavel-Dinu - Stanford UniversitySofia E. Luna - Stanford UniversityM. Kyle Cromer - Stanford UniversityRuhi Sayana - Stanford UniversityNatalia Gomez-Ospina - Stanford UniversityMatthew H. Porteus - Stanford University
- Resource Type
- Journal article
- Publication Details
- Nature biotechnology, Vol.42(5), pp.731-744
- DOI
- 10.1038/s41587-023-01888-4
- PMID
- 37537500
- NLM abbreviation
- Nat Biotechnol
- ISSN
- 1087-0156
- eISSN
- 1546-1696
- Publisher
- NATURE PORTFOLIO
- Number of pages
- 39
- Grant note
- Taube and Koret Foundation
- Language
- English
- Date published
- 05/01/2024
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984822991202771
Metrics
5 Record Views