CUT&Tag for efficient epigenomic profiling of small samples and single cells
CUT&Tag for efficient epigenomic profiling of small samples and single cells
复制标题
DOI：
10.1038/s41467-019-09982-5
复制
发表时间：
2019-04-29
影响因子：
16.6
通讯作者：
Henikoff, Steven
中科院分区：
文献类型：
--
作者：
Kaya-Okur, Hatice S.;Wu, Steven J.;Henikoff, Steven
许多染色质特征在调节基因表达中起着关键作用。对基因调控的完整理解需要在小样本细胞中以高分辨率绘制特定的染色质特征。在这里，我们描述了切割下的目标和标签化（CUT和标签），酶系策略，提供高效的高分辨率测序库分析不同的染色质组分。在CUT&Tag中，染色质蛋白被特异性抗体原位结合，然后将蛋白质A-Tn 5转座酶融合蛋白拴在一起。转座酶的激活有效地产生具有高分辨率和异常低背景的片段文库。从活细胞到测序就绪文库的所有步骤都可以在工作台上的单管或高通量管道中的微孔中进行，整个过程可以在一天内完成。我们通过分析低细胞数量和单细胞上的组蛋白修饰、RNA聚合酶II和转录因子来证明CUT和标签的实用性。
Many chromatin features play critical roles in regulating gene expression. A complete understanding of gene regulation will require the mapping of specific chromatin features in small samples of cells at high resolution. Here we describe Cleavage Under Targets and Tagmentation (CUT&Tag), an enzyme-tethering strategy that provides efficient high-resolution sequencing libraries for profiling diverse chromatin components. In CUT&Tag, a chromatin protein is bound in situ by a specific antibody, which then tethers a protein A-Tn5 transposase fusion protein. Activation of the transposase efficiently generates fragment libraries with high resolution and exceptionally low background. All steps from live cells to sequencing-ready libraries can be performed in a single tube on the benchtop or a microwell in a high-throughput pipeline, and the entire procedure can be performed in one day. We demonstrate the utility of CUT&Tag by profiling histone modifications, RNA Polymerase II and transcription factors on low cell numbers and single cells.