Genomic profiling of native R loops with a DNA-RNA hybrid recognition sensor.

Genomic profiling of native R loops with a DNA-RNA hybrid recognition sensor.
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利用一种DNA - RNA杂交识别传感器对天然R环进行基因组分析。

DOI:
10.1126/sciadv.abe3516
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发表时间:
2021-03
期刊:
影响因子:
13.6
通讯作者:
Liang K
Liang K
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Wang K;Wang H;Li C;Yin Z;Xiao R;Li Q;Xiang Y;Wang W;Huang J;Chen L;Fang P;Liang K

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DNA-RNA杂交识别传感器蛋白质在基因组中描绘天然R环结构。R环是一种独特的三链结构,参与多个关键生物过程,并与人类疾病相关。准确和全面的R环分析是R环研究的先决条件。然而,目前的R环映射方法产生很大的差异,因此,一个独立的方法是迫切需要的。在这里,我们建立了一个独立的R环CUT&Tag(Tn 5为基础的切割下的目标和标签化)的方法,通过结合CUT&Tag和GST-His 6 -2×HBD(谷胱甘肽S-转移酶-六组氨酸-2×杂交结合域),一个人工的DNA-RNA杂交传感器,特异性地识别DNA-RNA杂交。我们证明了R环切割和标签是敏感的,可重复的,方便的本地R环映射与高分辨率,并发现捕获策略,而不是传感器的特异性,在很大程度上有助于不同方法之间的差异。总之,我们提供了一个独立的策略,天然R环的基因组分析,并帮助解决多种R环作图方法之间的差异。
A DNA-RNA hybrid recognition sensor protein profiles native R loop structures in the genome. An R loop is a unique triple-stranded structure that participates in multiple key biological processes and is relevant to human diseases. Accurate and comprehensive R loop profiling is a prerequisite for R loops studies. However, current R loop mapping methods generate large discrepancies, therefore an independent method is in urgent need. Here, we establish an independent R loop CUT&Tag (Tn5-based cleavage under targets and tagmentation) method by combining CUT&Tag and GST-His6-2×HBD (glutathione S-transferase–hexahistidine–2× hybrid-binding domain), an artificial DNA-RNA hybrid sensor that specifically recognizes the DNA-RNA hybrids. We demonstrate that the R loop CUT&Tag is sensitive, reproducible, and convenient for native R loop mapping with high resolution, and find that the capture strategies, instead of the specificity of sensors, largely contribute to the disparities among different methods. Together, we provide an independent strategy for genomic profiling of native R loops and help resolve discrepancies among multiple R loop mapping methods.
DOI: 10.1016/j.cell.2018.09.027
发表时间: 2018-10-18
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影响因子: 64.5
作者:
Liang K;Smith ER;Aoi Y;Stoltz KL;Katagi H;Woodfin AR;Rendleman EJ;Marshall SA;Murray DC;Wang L;Ozark PA;Mishra RK;Hashizume R;Schiltz GE;Shilatifard A
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发表时间: 2020-06-01
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发表时间: 2020-08-20
影响因子: 14.9
作者:
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通讯作者: Cimprich KA
R环的形成是未甲基化的人类CpG岛启动子的独特特征。
DOI: 10.1016/j.molcel.2012.01.017
发表时间: 2012-03-30
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影响因子: 16
作者:
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通讯作者: Chédin F