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<p>Developing glycosylase-based base editors (gBEs) to broaden the editing scope is highly desirable for biomedical research and agricultural applications. However, the off-target effects and applicability of gBEs need further investigation. We employ GOTI to detect rare DNA off-target events in mouse embryos injected with N-methylpurine glycosylase-based AYBE and gGBE. Transcriptome-wide RNA analysis reveals that TadA8e-V106W, derived from AYBE, induces low-frequency RNA off-target editing. Both base editors efficiently induce A/G-to-Y editing in mouse and sheep embryos, and in newborn lambs. The robust efficiency and specificity of AYBE and gGBE underscore their potential for clinical applications and genetic improvement in livestock.</p>

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Efficient glycosylase-mediated base editing with minimal off-target effects in mammalian embryos

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月21日

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  • Yinghui Wei,
  • Xi Cao,
  • Shuhong Huang,
  • Yang Yue,
  • Muhua Luo,
  • Zujiang Liu,
  • Bo Liu,
  • Qijing Zhang,
  • Yangsheng Wu,
  • Liqin Wang,
  • Jiapeng Lin,
  • Yulin Chen,
  • Wenxin Zheng,
  • Weiwei Wu,
  • Kun Xu,
  • Xiaolong Wang

摘要

Developing glycosylase-based base editors (gBEs) to broaden the editing scope is highly desirable for biomedical research and agricultural applications. However, the off-target effects and applicability of gBEs need further investigation. We employ GOTI to detect rare DNA off-target events in mouse embryos injected with N-methylpurine glycosylase-based AYBE and gGBE. Transcriptome-wide RNA analysis reveals that TadA8e-V106W, derived from AYBE, induces low-frequency RNA off-target editing. Both base editors efficiently induce A/G-to-Y editing in mouse and sheep embryos, and in newborn lambs. The robust efficiency and specificity of AYBE and gGBE underscore their potential for clinical applications and genetic improvement in livestock.