Multimodal chromatin profiling using nanobody-based single-cell CUT&Tag.

Multimodal chromatin profiling using nanobody-based single-cell CUT&Tag.
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DOI:
10.1038/s41587-022-01535-4
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发表时间:
2023-06
影响因子:
46.9
通讯作者:
Castelo-Branco, Goncalo
Castelo-Branco, Goncalo
中科院分区:
工程技术1区
文献类型:
--
作者:
Bartosovic, Marek;Castelo-Branco, Goncalo

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通过单细胞CUT&Tag等技术,可以在数千个细胞中的单细胞水平上探测组蛋白修饰。在这里,我们描述了纳米切割和标签(纳米CT),它允许同时映射多达三个表观基因组模式在单细胞分辨率使用nanobody-Tn 5融合蛋白。多模式纳米CT与低至25,000 - 200,000个细胞的起始材料兼容,并且比单细胞CUT&Tag具有显著更高的灵敏度和每个细胞的碎片数量。我们使用nano-CT同时分析幼年小鼠脑中的染色质可及性,H3 K27 ac和H3 K27 me 3,从而比单峰单细胞CUT&Tag能够区分更多的细胞类型和状态。我们还推断了少突胶质细胞谱系中转座酶可及染色质(ATAC)和H3 K27 ac测定之间的染色质速度和去卷积H3 K27 me 3抑制状态,发现在少突胶质细胞谱系进展期间不同基因模块处的H3 K27 me 3抑制的两个连续波。鉴于其高分辨率,多功能性和多模式特征,nano-CT可以在单细胞水平上对复杂生物系统中的表观遗传景观进行独特的见解。使用Tn 5-纳米抗体融合蛋白以单细胞分辨率绘制三个表观遗传特征
Probing histone modifications at a single-cell level in thousands of cells has been enabled by technologies such as single-cell CUT&Tag. Here we describe nano-CUT&Tag (nano-CT), which allows simultaneous mapping of up to three epigenomic modalities at single-cell resolution using nanobody-Tn5 fusion proteins. Multimodal nano-CT is compatible with starting materials as low as 25,000–200,000 cells and has significantly higher sensitivity and number of fragments per cell than single-cell CUT&Tag. We use nano-CT to simultaneously profile chromatin accessibility, H3K27ac, and H3K27me3 in juvenile mouse brain, allowing for discrimination of more cell types and states than unimodal single-cell CUT&Tag. We also infer chromatin velocity between assay for transposase-accessible chromatin (ATAC) and H3K27ac in the oligodendrocyte lineage and deconvolute H3K27me3 repressive states, finding two sequential waves of H3K27me3 repression at distinct gene modules during oligodendrocyte lineage progression. Given its high resolution, versatility, and multimodal features, nano-CT allows unique insights in epigenetic landscapes in complex biological systems at the single-cell level. Three epigenetic features are mapped at single-cell resolution using Tn5-nanobody fusion proteins
DOI: 10.1083/jcb.201709115
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