Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)
Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)
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DOI：
10.1038/s41467-019-11559-1
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发表时间：
2019-08-20
影响因子：
16.6
通讯作者：
Zhao, Keji
中科院分区：
文献类型：
--
作者：
Carter, Benjamin;Ku, Wai Lim;Zhao, Keji
现代下一代测序方法使研究人员能够分析单个细胞的表观遗传状态。用于在单细胞中分析表观遗传标记的现有技术通常需要使用和优化时间密集型程序，例如液滴射流、染色质片段化和末端修复。在这里，我们描述了ACT-seq，一种用于在少量或单个细胞中绘制组蛋白尾部修饰、组蛋白变体和染色质结合蛋白的全基因组分布的简化方法。ACT-seq利用Tn 5转座酶与蛋白A的融合，蛋白A通过特异性抗体靶向染色质，允许染色质片段化和序列标签特异性插入呈递相关抗原的基因组位点。Tn 5转座酶使得能够使用索引多路复用策略（iACT-seq），其使得能够由单个研究人员在一天内构建数千个单细胞文库，而不需要基于液滴的流体学或视觉分选。我们的结论是，ACT-seq提出了一个有吸引力的替代现有的技术在单细胞中的表观遗传标记。
Modern next-generation sequencing-based methods have empowered researchers to assay the epigenetic states of individual cells. Existing techniques for profiling epigenetic marks in single cells often require the use and optimization of time-intensive procedures such as drop fluidics, chromatin fragmentation, and end repair. Here we describe ACT-seq, a streamlined method for mapping genome-wide distributions of histone tail modifications, histone variants, and chromatin-binding proteins in a small number of or single cells. ACT-seq utilizes a fusion of Tn5 transposase to Protein A that is targeted to chromatin by a specific antibody, allowing chromatin fragmentation and sequence tag insertion specifically at genomic sites presenting the relevant antigen. The Tn5 transposase enables the use of an index multiplexing strategy (iACT-seq), which enables construction of thousands of single-cell libraries in one day by a single researcher without the need for drop-based fluidics or visual sorting. We conclude that ACT-seq present an attractive alternative to existing techniques for mapping epigenetic marks in single cells.