2020年7月27日上线的《自然—方法学》杂志发表了美国科学家的一项最新研究成果。哈佛医学院的C.-ting Wu课题组利用原位测序实现了对人类基因组的三维映射和加速超分辨率成像。

该课题组人员引入OligoFISSEQ，提出三种方式将荧光原位测序（FISSEQ）编码Oligopaint探针用于实现许多定向的基因组区域的快速可视化。将OligoFISSEQ应用于人类二倍体成纤维细胞，该团队展示了利用四轮测序就足以产生成百上千个细胞的、分布在6个染色体上36个基因组目标的三维映射。这意味OligoFISSEQ具有在5-8轮测序下成像几千个基因组目标的潜力。

研究组还用OligoFISSEQ方法在更高分辨率下追踪染色体，揭示了X染色体穿过46个区域的路径。OligoFISSEQ也与免疫细胞化学技术具有相容性。最后，研究人员将OligoFISSEQ与OligoSTORM结合，奠定了对人类大多数基因组（或全基因组）的加速单分子超分辨率成像的基础。

据了解，对全基因组水平的染色体成像方法、以及更大的基因组覆盖和更高的光学分辨率仍有需求。

附：英文原文

Title: 3D mapping and accelerated super-resolution imaging of the human genome using in situ sequencing

Author: Huy Q. Nguyen, Shyamtanu Chattoraj, David Castillo, Son C. Nguyen, Guy Nir, Antonios Lioutas, Elliot A. Hershberg, Nuno M. C. Martins, Paul L. Reginato, Mohammed Hannan, Brian J. Beliveau, George M. Church, Evan R. Daugharthy, Marc A. Marti-Renom, C.-ting Wu

Issue&Volume: 2020-07-27

Abstract: There is a need for methods that can image chromosomes with genome-wide coverage, as well as greater genomic and optical resolution. We introduce OligoFISSEQ, a suite of three methods that leverage fluorescence in situ sequencing (FISSEQ) of barcoded Oligopaint probes to enable the rapid visualization of many targeted genomic regions. Applying OligoFISSEQ to human diploid fibroblast cells, we show how four rounds of sequencing are sufficient to produce 3D maps of 36 genomic targets across six chromosomes in hundreds to thousands of cells, implying a potential to image thousands of targets in only five to eight rounds of sequencing. We also use OligoFISSEQ to trace chromosomes at finer resolution, following the path of the X chromosome through 46 regions, with separate studies showing compatibility of OligoFISSEQ with immunocytochemistry. Finally, we combined OligoFISSEQ with OligoSTORM, laying the foundation for accelerated single-molecule super-resolution imaging of large swaths of, if not entire, human genomes.

DOI: 10.1038/s41592-020-0890-0

Source: https://www.nature.com/articles/s41592-020-0890-0