Undetectable off-target effects induced by FokI catalytic domain in mouse embryos
2024
Long Xie | Hu Feng | Zhifang Li | Di Li | Xiali Yang | Tanglong Yuan | Nana Yan | Chenfei He | Jitan Zheng | Zhenrui Zuo | Yaxuan Zheng | Yaqi Cao | Yangqing Lu | Xing Yao Xiong | Erwei Zuo
Abstract The FokI catalytic domain can be fused to various DNA binding architectures to improve the precision of genome editing tools. However, evaluation of off-target effects is essential for developing these tools. We use Genome-wide Off-target analysis by Two-cell embryo Injection (GOTI) to detect low-frequency off-target editing events in mouse embryos injected with FokI-based architectures. Specifically, we test FokI-heterodimers fused with TALENs, FokI homodimers fused with RYdCas9, or FokI catalytic domains alone resulting in no significant off-target effects. These FokI genome editing systems exhibit undetectable off-target effects in mouse embryos, supporting the further development of these systems for clinical applications.
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