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

Area of Science:

• Virology
• Molecular Biology
• Nanotechnology

Background:

• Single virus tracking is crucial for understanding viral infection mechanisms.
• Efficiently labeling internal viral components without altering viral structure is challenging.
• Existing labeling strategies have limited applicability across diverse viruses.

Purpose of the Study:

• To develop a novel strategy for labeling viral nucleic acids with quantum dots (QDs) for real-time virus tracking.
• To apply the clustered regularly interspaced short palindromic repeats (CRISPR) imaging system for in situ viral labeling.
• To monitor the infection processes of Pseudorabies virus (PRV) using QD-labeled nucleic acids.

Main Methods:

• Conjugation of QDs to viral nucleic acids using nuclease-deactivated Cas9/gRNA complexes within living cells.
• Packaging of QD-conjugated viral nucleic acids into PRV virions during assembly.
• Real-time monitoring of PRV-QD adsorption, cytoplasmic transport, and nuclear entry in Vero and HeLa cells.

Main Results:

• Successful labeling of PRV nucleic acids with QDs was achieved.
• The CRISPR-based strategy enabled visualization of PRV infection dynamics.
• The method demonstrated efficient tracking of viral adsorption, transport, and nuclear entry.

Conclusions:

• The developed CRISPR imaging strategy offers an efficient and versatile method for labeling viral nucleic acids.
• This technique facilitates real-time studies of viral infection processes.
• The approach holds promise for advancing our understanding of virus-host interactions and developing antiviral strategies.