Abstract
Iatrogenic adverse events in clinical trials of retroviral vector-mediated gene-corrected cells have prioritized the urgent need for more comprehensive and stringent assessment of potentially genotoxic off-target alterations and the biosafety of cells intended for therapeutic applications. Genome editing tools such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced palindromic repeats (CRISPR)-Cas9 nuclease systems are being investigated as safer and efficient alternatives for site-directed genome modification. Using site-specific integration into the AAVS1 locus of primary human cells as an example, we present an integrated approach to multimodal investigation of off-target alterations and an evaluation of potential genotoxicity induced by ZFN-mediated integration of a therapeutic transgene.
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Acknowledgments
Supported by the National Medical Research Council (grant CIRG/1326/2012), National Cancer Centre and CellResearch Corporation (all in Singapore). D.K. and S.M.S. were supported by the Agency for Science, Technology and Research (A*STAR), Singapore.
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Sivalingam, J. et al. (2018). Integrated Multimodal Evaluation of Genotoxicity in ZFN-Modified Primary Human Cells. In: Liu, J. (eds) Zinc Finger Proteins. Methods in Molecular Biology, vol 1867. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8799-3_11
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DOI: https://doi.org/10.1007/978-1-4939-8799-3_11
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