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纳米-CUT&Tag用于单细胞分辨率的多式染色体分析

José Ramón Bárcenas-Walls¹, Federico Ansaloni², Bastien Hervé²

¹Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.

Nature protocols

|December 22, 2023

概括

我们开发了纳米CT,这是一种用于单细胞表观遗传分析的新方法. 这种技术同时分析基因素标记和开放色素,进步我们对复杂生物系统中基因调节的理解.

科学领域:

分子生物学分子生物学
表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.
基因组学就是基因组学.

背景情况:

了解复杂组织中的基因调节需要对染色质状态进行单细胞分辨率分析.
同时对多个表观遗传标记进行分析对于全面分析至关重要.

研究的目的:

引入和验证一种基于纳米体的新型单细胞CUT&Tag (纳米-CT) 协议.
从单个细胞中实现三种表观遗传模式 (两个基因组标记和开放色素) 的同时分析.

主要方法:

开发新的二次纳米体-Tn5融合蛋白,用于有针对性的条形码标记.
使用商用平台进行单细胞索引.
纳米体-Tn5融合蛋白的内部生产和净化 (3-4天).

主要成果:

纳米CT成功地产生了来自数千个脑细胞的多模式表观基因组资料,以单细胞分辨率.
该协议是高效的,需要很少的样本输入,并且在模式选择中提供灵活性.
公司内部生产必不可少的试剂在3-4天内是可行的.

结论:

纳米-CT为多式单细胞表观基因组分析提供了一个强大,易于使用的工具.
该方法有助于研究表观遗传异质性和发育和疾病中的染色质动态.
有潜力揭示不同染色质模式之间的相互作用,以获得更深入的生物学见解.

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