Identification of Transcription Factor Binding Sites by Cleavage Under Target and Release Using Nuclease in Zebrafish.

Identification of Transcription Factor Binding Sites by Cleavage Under Target and Release Using Nuclease in Zebrafish.
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DOI:
10.1089/zeb.2021.0082
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发表时间:
2022-06
期刊:
影响因子:
2
通讯作者:
Espin-Palazon, Raquel
Espin-Palazon, Raquel
中科院分区:
生物学4区
文献类型:
--
作者:
Barakat, Radwa;Campbell, Clyde A.;Espin-Palazon, Raquel

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靶下切割和使用核酸酶释放(CUT&RUN)已经成为优于传统方法的染色质分析策略。虽然CUT&RUN已被广泛用于哺乳动物细胞,但其在斑马鱼中的应用仍处于早期阶段。在这项研究中,我们已经开发了一个协议,成功地进行切割和运行映射转录因子(TF)结合位点在胚胎,成体组织,和流式细胞仪分选的斑马鱼细胞。我们还提供了一个详细的工作流程,用于识别预测TF结合位点,可以在任何动物物种中使用。总而言之,我们的策略将把这一宝贵的工具扩展到斑马鱼社区,提高在这种模式生物中可以实现的表观遗传分辨率。
Cleavage Under Targets and Release Using Nuclease (CUT&RUN) has emerged as a chromatin profiling strategy that excels traditional methods. Although CUT&RUN has been widely utilized in mammalian cells, its use in the zebrafish is at its early stages. In this study, we have developed a protocol to successfully perform CUT&RUN to map transcription factor (TF) binding sites in embryonic, adult tissues, and FACS-sorted zebrafish cells. We also provide a detailed workflow for the identification of predicted TF binding sites that can be utilized in any animal species. Altogether, our strategy will expand this invaluable tool to the zebrafish community, improving the epigenetic resolution that can be achieved in this model organism.
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