Dam Assisted Fluorescent Tagging of Chromatin Accessibility (DAFCA) for Optical Genome Mapping in Nanochannel Arrays.

Dam Assisted Fluorescent Tagging of Chromatin Accessibility (DAFCA) for Optical Genome Mapping in Nanochannel Arrays.
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DOI:
10.1021/acsnano.2c12755
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发表时间:
2023-05-23
期刊:
影响因子:
17.1
通讯作者:
Ebenstein, Yuval
Ebenstein, Yuval
中科院分区:
材料科学1区
文献类型:
--
作者:
Nifker, Gil;Grunwald, Assaf;Margalit, Sapir;Tulpova, Zuzana;Michaeli, Yael;Har-Gil, Hagai;Maimon, Noy;Roichman, Elad;Schuetz, Leonie;Weinhold, Elmar;Ebenstein, Yuval

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细胞核中的蛋白质和酶需要物理访问其DNA靶位点,以执行基因组任务,如基因激活和转录。因此,染色质可及性是基因表达的中心调节因子,其基因组图谱包含关于细胞类型和状态的重要信息。我们使用了E。大肠杆菌Dam甲基转移酶与荧光辅因子类似物的组合,以在细胞核内可接近的DNA区域中产生荧光标签。然后通过纳米通道阵列中的单分子光学基因组作图来检测基因组的可接近部分。这种方法使我们能够表征长程结构变异及其相关的染色质结构。我们展示了创建由在硅纳米通道中延伸的长DNA分子组成的全基因组、等位基因特异性染色质可及性图的能力。
Proteins and enzymes in the cell nucleus require physical access to their DNA target sites in order to perform genomic tasks such as gene activation and transcription. Hence, chromatin accessibility is a central regulator of gene expression, and its genomic profile holds essential information on the cell type and state. We utilized the E. coli Dam methyltransferase in combination with a fluorescent cofactor analogue to generate fluorescent tags in accessible DNA regions within the cell nucleus. The accessible portions of the genome are then detected by single-molecule optical genome mapping in nanochannel arrays. This method allowed us to characterize long-range structural variations and their associated chromatin structure. We show the ability to create whole-genome, allele-specific chromatin accessibility maps composed of long DNA molecules extended in silicon nanochannels.
全基因组癌症特异性染色质的可及性模式来自档案处理的异种移植肿瘤。
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