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Developmental Biology
在小鼠成肌细胞研究中使用靶标和标记下的切割(CUT&Tag)测定
在小鼠成肌细胞研究中使用靶标和标记下的切割(CUT&Tag)测定
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Developmental Biology
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JoVE Journal Developmental Biology
Using Cleavage Under Targets and Tagmentation (CUT&Tag) Assay in Mouse Myoblast Research

在小鼠成肌细胞研究中使用靶标和标记下的切割(CUT&Tag)测定

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1,306 Views
07:09 min
March 1, 2024

DOI: 10.3791/66066-v

Yuefeng Li1,2, Xiaofen Wu1,2, Ping Hu1,2

1Guangzhou Laboratory, 2The Key Laboratory of Biological Targeting Diagnosis, Therapy and Rehabilitation of Guangdong Higher Education Institutes,The Fifth Affiliated Hospital of Guangzhou Medical University

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刚进入表观遗传学领域的研究人员会发现 CUT&Tag 是 ChIP 检测的明显更容易的替代品。CUT&Tag极大地促进了稀有细胞群和原代细胞群的表观遗传学研究,从极少数细胞中生成了高质量的数据。该协议描述了在从小鼠后肢肌肉分离的小鼠成肌细胞上进行H3K4me1 CUT和Tag测定。

我们小组的研究重点是成人骨骼肌干细胞的基因组结构和转录如何受到调节和维持,特别是在硫酸盐变化期间,如干细胞承诺、分化和滑痹。成体骨骼肌干细胞在肌肉组织中仅占单核细胞的 1%,由于它们在从肌肉消化中获得的单细胞悬浮液中的稀缺性,因此对使用 High-C、ChIP 和 ATAC seq 的染色质特征研究提出了挑战。染色质免疫沉淀 (ChIP) 一直是分析基因组特征的关键工具。

Steven Henikoff 博士开发的一种更简单、更高效的切割和标签技术通常超过 ChIP,尤其是在研究稀有细胞群的表观遗传学方面。它的易用性扩大了研究人员(包括非表观遗传学专家)的可访问性。

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关键词:CUT&Tag 靶标和标记下的裂解 小鼠成肌细胞 肌肉干细胞 染色质结合因子 组蛋白标记 组蛋白变体 ChIP 染色质免疫沉淀 交联 ChIP 天然 ChIP 微球菌核酸酶 测序 基因组位置

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