Integrated Multimodal Evaluation of Genotoxicity in ZFN-Modified Primary Human Cells

• DOI: 10.1007/978-1-4939-8799-3_11
• Publication Date: 2018-08-28T01:02:37Z
• Authors: Jaichandran Sival...; Dimitar Kenanov; Wai Har Ng; Sze Sing Lee; Toan Thang Phan; Sebastian Maurer-...; Oi Lian Kon
• Keywords: Zinc finger nuclease; Nuclease

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.