Single Virus Tracking with Quantum Dots Packaged into Enveloped Viruses Using CRISPR

Labeling viruses with high-photoluminescence quantum dots (QDs) for single virus tracking provides a visual tool to aid our understanding of viral infection mechanisms. However, efficiently labeling internal viral components without modifying the viral envelope and capsid remains a challenge, and ex...

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Bibliographic Details
Published in:Nano letters 2020-02, Vol.20 (2), p.1417-1427
Main Authors: Yang, Yong-Bo, Tang, Yan-Dong, Hu, Yue, Yu, Fang, Xiong, Jun-Yao, Sun, Ming-Xia, Lyu, Chuang, Peng, Jin-Mei, Tian, Zhi-Jun, Cai, Xue-Hui, An, Tong-Qing
Format: Article
Language:English
Subjects:
Capsid
CRISPR-Cas Systems - genetics
HeLa Cells
Herpesvirus 1, Suid - isolation & purification
Herpesvirus 1, Suid - ultrastructure
Humans
Quantum Dots - chemistry
Virion - genetics
Virion - isolation & purification
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Summary:Labeling viruses with high-photoluminescence quantum dots (QDs) for single virus tracking provides a visual tool to aid our understanding of viral infection mechanisms. However, efficiently labeling internal viral components without modifying the viral envelope and capsid remains a challenge, and existing strategies are not applicable to most viruses. Here, we have devised a strategy using the clustered regularly interspaced short palindromic repeats (CRISPR) imaging system to label the nucleic acids of Pseudorabies virus (PRV) with QDs. In this strategy, QDs were conjugated to viral nucleic acids with the help of nuclease-deactivated Cas9/gRNA complexes in the nuclei of living cells and then packaged into PRV during virion assembly. The processes of PRV-QD adsorption, cytoplasmic transport along microtubules, and nuclear entry were monitored in real time in both Vero and HeLa cells, demonstrating the utility and efficiency of the strategy in the study of viral infection.
ISSN:1530-6984
1530-6992
DOI:10.1021/acs.nanolett.9b05103