3D mapping and accelerated super-resolution imaging of the human genome using in situ sequencing.
3D mapping and accelerated super-resolution imaging of the human genome using in situ sequencing.
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DOI：
10.1038/s41592-020-0890-0
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发表时间：
2020-08
期刊：
影响因子：
48
通讯作者：
Wu CT
中科院分区：
文献类型：
--
作者：
Nguyen HQ;Chattoraj S;Castillo D;Nguyen SC;Nir G;Lioutas A;Hershberg EA;Martins NMC;Reginato PL;Hannan M;Beliveau BJ;Church GM;Daugharthy ER;Marti-Renom MA;Wu CT
基因组功能可能会对其三维(3D)组织做出反应，这突显了对能够以更高的覆盖率以及更高的基因组和光学分辨率来成像基因组的方法的需求。在这里，我们通过引入OglioFISSEQ来推动这一目标，这是一套三种方法，利用条形码寡核苷酸探针的荧光原位测序来实现许多目标基因组区域的快速可视化。将寡聚FISSEQ应用于人类二倍体成纤维细胞，我们展示了只需四轮测序就足以在数百到数千个细胞中产生跨越6条染色体的66个基因组目标的3D图谱。然后，我们使用寡聚FISSEQ以更高的分辨率追踪染色体，沿着X染色体的路径穿过46个区域，不同的研究表明，寡聚FISSEQ与免疫化学是兼容的。最后，我们将寡聚FISSEQ和寡聚STORM结合在一起，为加速对大片(如果不是整个)人类基因组进行单分子超分辨率成像奠定了基础。
That genome function may respond to its three-dimensional (3D) organization highlights the need for methods that can image genomes with superior coverage as well as greater genomic and optical resolution. Here, we push toward this goal by introducing OligoFISSEQ, a suite of three methods that leverage fluorescent in situ sequencing of barcoded Oligopaint probes to enable the rapid visualization of many targeted genomic regions. Applying OligoFISSEQ to human diploid fibroblast cells, we show how only four rounds of sequencing are sufficient to produce 3D maps of 66 genomic targets across 6 chromosomes in hundreds to thousands of cells. We then 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 immunochemistry. 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.
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光学成像。膨胀显微镜。
DOI：
10.1126/science.1260088
发表时间：
2015-01-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen F;Tillberg PW;Boyden ES
通讯作者：
Boyden ES
影响因子：
16.6
作者：
Cattoni DI;Cardozo Gizzi AM;Georgieva M;Di Stefano M;Valeri A;Chamousset D;Houbron C;Déjardin S;Fiche JB;González I;Chang JM;Sexton T;Marti-Renom MA;Bantignies F;Cavalli G;Nollmann M
通讯作者：
Nollmann M
影响因子：
4
作者：
Joyce EF;Erceg J;Wu CT
通讯作者：
Wu CT
影响因子：
16
作者：
Cardozo Gizzi, Andres M.;Cattoni, Diego, I;Nollmann, Marcelo
通讯作者：
Nollmann, Marcelo
影响因子：
12.3
作者：
Deng X;Ma W;Ramani V;Hill A;Yang F;Ay F;Berletch JB;Blau CA;Shendure J;Duan Z;Noble WS;Disteche CM
通讯作者：
Disteche CM