生物
基因组
人类基因组
计算生物学
基因组学
染色体
荧光原位杂交
遗传学
基因
作者
Huy Q. Nguyen,Shyamtanu Chattoraj,David Castillo,Son C. Nguyen,Guy Nir,Antonios Lioutas,Elliot A. Hershberg,Nuno M. C. Martins,Paul Reginato,Mohammed A. Hannan,Brian J. Beliveau,George M. Church,Evan R Daugharthy,Marc A. Martí‐Renom,Chao-ting Wu
出处
期刊:Nature Methods
[Springer Nature]
日期:2020-07-27
卷期号:17 (8): 822-832
被引量:112
标识
DOI:10.1038/s41592-020-0890-0
摘要
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. OligoFISSEQ combines Oligopaints with fluorescence in situ sequencing to enable the 3D mapping of many regions across the genome in human cells to interrogate genome organization at improved genomic resolution. OligoFISSEQ is compatible with immunochemistry and OligoSTORM for super-resolution imaging.
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