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Zygotic genome activation (ZGA) triggered by transcription factors (TFs) is a critical event to facilitate subsequent embryonic development. However, the paucity of material poses a challenge for mapping TF binding profiles in mammalian embryos. Here, we describe a low-input Cleavage Under Targets & Tagmentation (CUT&Tag) protocol optimized for TF binding using ~300 mouse two-cell embryos to investigate ZGA. This version of CUT&Tag is an immunostaining-based protocol with manual cell transfer by pipette to avoid loss of samples and gentle washing of fragile preimplantation embryos protected by the zona pellucida.

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CUT&Tag for Transcription Factor Binding Profiles During Mammalian Zygotic Genome Activation

  • 实验方案

  • 发表日期：

    2025年

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  • Wataru Kobayashi,
  • Kikuë Tachibana

摘要

Zygotic genome activation (ZGA) triggered by transcription factors (TFs) is a critical event to facilitate subsequent embryonic development. However, the paucity of material poses a challenge for mapping TF binding profiles in mammalian embryos. Here, we describe a low-input Cleavage Under Targets & Tagmentation (CUT&Tag) protocol optimized for TF binding using ~300 mouse two-cell embryos to investigate ZGA. This version of CUT&Tag is an immunostaining-based protocol with manual cell transfer by pipette to avoid loss of samples and gentle washing of fragile preimplantation embryos protected by the zona pellucida.