Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Key Laboratory of Synthetic Biology, Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science and Technology, Guangxi University, Nanning, 530004, China.
Genome Biol. 2024 Feb 20;25(1):51. doi: 10.1186/s13059-024-03188-9.
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.
FokI 催化结构域可以与各种 DNA 结合结构融合，以提高基因组编辑工具的精确性。然而，评估脱靶效应对于开发这些工具至关重要。我们使用通过双细胞胚胎注射的全基因组脱靶分析（GOTI）来检测用 FokI 为基础的结构注射到小鼠胚胎中的低频脱靶编辑事件。具体来说，我们测试了与 TALENs 融合的 FokI 杂二聚体、与 RYdCas9 融合的 FokI 同二聚体，或单独的 FokI 催化结构域，结果均未显示出明显的脱靶效应。这些 FokI 基因组编辑系统在小鼠胚胎中未显示出可检测的脱靶效应，支持进一步将这些系统应用于临床。
