Live Visualization of HIV-1 Proviral DNA Using a Dual-Color-Labeled CRISPR System,Analytical Chemistry

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Live Visualization of HIV-1 Proviral DNA Using a Dual-Color-Labeled CRISPR System
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-21 00:00:00 , DOI: 10.1021/acs.analchem.7b03584
Yingxin Ma _{1,

2} , Mingxiu Wang ₁ , Wei Li ₁ , Zhiping Zhang ₁ , Xiaowei Zhang ₁ , Guoqiang Wu ₁ , Tianwei Tan ₂ , Zongqiang Cui ₁ , Xian-En Zhang ₃

Affiliation

HIV latency is one of the major problems in HIV/AIDS cure. Imaging single-copy integrated proviral HIV DNA in host cell has both virology and clinical significance but remains technical challenge. Here, we developed a dual-color labeled CRISPR system to image the HIV-1 integrated proviral DNA in latently infected cells. The pair of CRISPRs was fluorescently labeled with two different color QDs using two alternative bioorthogonal ligation reactions. Integrated HIV-sequences are successfully mapped based on the colocalized signals of QDs in living cells. Compared to the existing zinc finger proteins and TALENs, the CRISPR system is much easier to operate and more efficient in imaging of internal genomic loci. Therefore, the proposed method could be not only a powerful tool for imaging proviral HIV-1, but also a versatile platform to image single genomic loci in living cells.

中文翻译：

使用双色标签CRISPR系统对HIV-1前病毒DNA进行实时可视化

艾滋病毒潜伏期是艾滋病毒/艾滋病治疗的主要问题之一。在宿主细胞中对单拷贝整合的原病毒HIV DNA进行成像具有病毒学和临床意义，但仍然是技术挑战。在这里，我们开发了一种双色标记的CRISPR系统，可对潜伏感染细胞中HIV-1整合的原病毒DNA进行成像。使用两个不同的生物正交连接反应，用两个不同颜色的QD对这对CRISPRs进行荧光标记。基于活细胞中QD的共定位信号，成功绘制了整合的HIV序列。与现有的锌指蛋白和TALENs相比，CRISPR系统更易于操作，并且在内部基因组位点的成像方面更为有效。因此，提出的方法不仅可以作为对原病毒HIV-1进行成像的强大工具，

更新日期：2017-11-21

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