Single Virus Tracking with Quantum Dots Packaged into Enveloped Viruses Using CRISPR.
Nano Lett
; 20(2): 1417-1427, 2020 02 12.
Article
em En
| MEDLINE
| ID: mdl-31930919
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírion
/
Herpesvirus Suídeo 1
/
Pontos Quânticos
/
Sistemas CRISPR-Cas
Limite:
Humans
Idioma:
En
Revista:
Nano Lett
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
China