DNA nanomapping using CRISPR-Cas9 as a programmable nanoparticle.

DNA nanomapping using CRISPR-Cas9 as a programmable nanoparticle.
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DOI:
10.1038/s41467-017-01891-9
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发表时间:
2017-11-21
影响因子:
16.6
通讯作者:
Reed J
Reed J
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Mikheikin A;Olsen A;Leslie K;Russell-Pavier F;Yacoot A;Picco L;Payton O;Toor A;Chesney A;Gimzewski JK;Mishra B;Reed J

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使用短读段的全基因组测序的进展(例如,<150 bp),下一代测序技术重新激发了人们对高分辨率物理作图的兴趣,以填补测序无法很好解决的技术空白。在这里,我们报告了DNA纳米技术和单分子基因组学的两项技术进展:(1)我们描述了一种标记技术(CRISPR-Cas9纳米颗粒),用于基于高速AFM的DNA物理映射;(2)首次成功演示了使用DVD光学器件通过高速AFM对DNA分子进行成像。作为原理的证明,我们使用这种新的“纳米绘图”方法来精确检测和绘制滤泡性淋巴瘤患者淋巴结活检中存在的BCL 2-IGH易位。这种HS-AFM“纳米绘图”技术可以与测序和其他物理绘图方法互补。DNA的物理作图可用于检测结构变体和用于全基因组单倍型组装。在这里,作者使用CRISPR-Cas9和高速原子力显微镜来“纳米映射”DNA的单个分子。
Progress in whole-genome sequencing using short-read (e.g., <150 bp), next-generation sequencing technologies has reinvigorated interest in high-resolution physical mapping to fill technical gaps that are not well addressed by sequencing. Here, we report two technical advances in DNA nanotechnology and single-molecule genomics: (1) we describe a labeling technique (CRISPR-Cas9 nanoparticles) for high-speed AFM-based physical mapping of DNA and (2) the first successful demonstration of using DVD optics to image DNA molecules with high-speed AFM. As a proof of principle, we used this new “nanomapping” method to detect and map precisely BCL2–IGH translocations present in lymph node biopsies of follicular lymphoma patents. This HS-AFM “nanomapping” technique can be complementary to both sequencing and other physical mapping approaches. Physical mapping of DNA can be used to detect structural variants and for whole-genome haplotype assembly. Here, the authors use CRISPR-Cas9 and high-speed atomic force microscopy to ‘nanomap’ single molecules of DNA.
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