CUT&Tag2for1: a modified method for simultaneous profiling of the accessible and silenced regulome in single cells.

CUT&Tag2for1: a modified method for simultaneous profiling of the accessible and silenced regulome in single cells.
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CUT&Tag2for1:一种改进的方法,用于同时分析单细胞中可访问和沉默的规则组。

DOI:
10.1186/s13059-022-02642-w
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发表时间:
2022-03-17
期刊:
影响因子:
12.3
通讯作者:
Henikoff S
Henikoff S
中科院分区:
生物学1区
文献类型:
--
作者:
Janssens DH;Otto DJ;Meers MP;Setty M;Ahmad K;Henikoff S

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靶下切割和标记(CUT&Tag)是一种抗体导向的转座酶系固策略,用于小样本和单细胞的原位染色质谱分析。我们描述了一种改进的CUT&Tag方案,使用RNA聚合酶II起始形式的抗体(Pol2丝氨酸-5磷酸)和抑制多梳结构域的抗体(H3K27me3)的混合物,然后通过计算信号反卷积生成单细胞中活性和抑制规则体的高分辨率地图。使用单一工作流程无缝绘制活性启动子、增强子和抑制调控元件的能力提供了适用于高通量单细胞平台的完整规则组分析策略。在线版本包含补充材料,下载地址:10.1186/s13059-022-02642-w。
Cleavage Under Targets and Tagmentation (CUT&Tag) is an antibody-directed transposase tethering strategy for in situ chromatin profiling in small samples and single cells. We describe a modified CUT&Tag protocol using a mixture of an antibody to the initiation form of RNA polymerase II (Pol2 Serine-5 phosphate) and an antibody to repressive Polycomb domains (H3K27me3) followed by computational signal deconvolution to produce high-resolution maps of both the active and repressive regulomes in single cells. The ability to seamlessly map active promoters, enhancers, and repressive regulatory elements using a single workflow provides a complete regulome profiling strategy suitable for high-throughput single-cell platforms. The online version contains supplementary material available at 10.1186/s13059-022-02642-w.
DOI: 10.21769/bioprotoc.4043
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影响因子: 0.8
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