优化的 CUT&Tag 能够在 Schizosaccharomyces pombe 中进行强大的表观基因组分析
Cheng-Zhi Huang1, Kang-Di Zhou1, Wei Ma1
1Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China; School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Methods (San Diego, Calif.)
|August 24, 2025
概括
我们优化了裂变酵母的透性和分片化 (CUT&Tag),使H3K9me3能够在最小的细胞输入下进行强大的分析. 这种新方法克服了高分辨率表观基因组分析的ChIP-seq限制.
科学领域:
- 表观遗传学
- 分子生物学
- 微生物学
背景情况:
- 染色体免疫沉测序 (ChIP-seq) 受到交叉连接器件和高细胞输入要求的限制.
- 需要优化高分辨率分析的方法来分析schizosaccharomyces pombe (S. pombe) 的表观基因组.
研究的目的:
- 优化S. pombe中CUT&Tag的透性,以便进行强大的H3K9me3分析.
- 建立一个高分辨率的表观基因组分析协议,减少细胞输入.
主要方法:
- 使用Lywallzyme优化S. pombe球状细胞生成 (>95%的效率).
- 系统性优化CUT&Tag的参数,包括酶度和细胞负载.
- 使用优化协议对野生型和clr4Δ菌株进行H3K9me3映射.
主要成果:
- 在形成球状细胞方面,Lywallzyme的性能优于其他酶.
- 确定了最佳的消化动力学和反向的细胞负载效率关系.
- 在信号与噪声比较高的中间体/端粒中捕获了特定的异色丰富.
结论:
- 优化的CUT&Tag协议使S. pombe在最小的细胞输入下进行了强大的表观遗传学分析.
- 这种方法克服了ChIP-seq的局限性,为表观遗传学研究提供了更好的分辨率.
相关概念视频
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01:46Epigenetic Regulation
Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
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