美国霍华德休斯医学研究所Zhe Liu、加州大学伯克利分校Robert Tjian等研究人员合作，开发了用于可及性基因组超分辨成像的新型显微镜。这一研究成果于2020年3月16日在线发表在《自然—方法学》上。

为了在纳米尺度上对可及性基因组进行原位成像，研究人员开发了转座酶可及性染色质光激活定位显微镜（3D ATAC-PALM），其将转座酶可及性染色质测定与可视化、PALM超分辨率成像以及晶格光片显微镜整合在一起。

3D ATAC-PALM与oligopaint DNA-荧光原位杂交（FISH）、RNA-FISH和蛋白质荧光多路复用，连接了显微镜和基因组数据，从而揭示了空间分隔的可及染色质域（ACD），其中包含活性染色质和转录的基因。

使用这些方法来分析遗传扰动的细胞，研究人员证明了基因组结构蛋白CTCF可以防止可及性染色质过度聚集并破坏ACD。

这些结果突显出3D ATAC-PALM是一种有用的工具，可探测基因组的结构和组织机制。

附：英文原文

Title: 3D ATAC-PALM: super-resolution imaging of the accessible genome

Author: Liangqi Xie, Peng Dong, Xingqi Chen, Tsung-Han S. Hsieh, Sambashiva Banala, Margherita De Marzio, Brian P. English, Yifeng Qi, Seol Kyoung Jung, Kyong-Rim Kieffer-Kwon, Wesley R. Legant, Anders S. Hansen, Anton Schulmann, Rafael Casellas, Bin Zhang, Eric Betzig, Luke D. Lavis, Howard Y. Chang, Robert Tjian, Zhe Liu

Issue&Volume: 2020-03-16

Abstract: To image the accessible genome at nanometer scale in situ, we developed three-dimensional assay for transposase-accessible chromatin-photoactivated localization microscopy (3D ATAC-PALM) that integrates an assay for transposase-accessible chromatin with visualization, PALM super-resolution imaging and lattice light-sheet microscopy. Multiplexed with oligopaint DNA–fluorescence in situ hybridization (FISH), RNA–FISH and protein fluorescence, 3D ATAC-PALM connected microscopy and genomic data, revealing spatially segregated accessible chromatin domains (ACDs) that enclose active chromatin and transcribed genes. Using these methods to analyze genetically perturbed cells, we demonstrated that genome architectural protein CTCF prevents excessive clustering of accessible chromatin and decompacts ACDs. These results highlight 3D ATAC-PALM as a useful tool to probe the structure and organizing mechanism of the genome.

DOI: 10.1038/s41592-020-0775-2

Source: https://www.nature.com/articles/s41592-020-0775-2