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Delaying Photobleaching of a Light-Switch Complex for Real-Time Imaging of Single Viral Particle Uncoating.

Delaying Photobleaching of a Light-Switch Complex for Real-Time Imaging of Single Viral Particle Uncoating.
Author Information (click to view)

Ma Y, Cui Z, He Z, Li W, Zhang ZP, Zhang X, Zhang XE, Tan T,


Ma Y, Cui Z, He Z, Li W, Zhang ZP, Zhang X, Zhang XE, Tan T, (click to view)

Ma Y, Cui Z, He Z, Li W, Zhang ZP, Zhang X, Zhang XE, Tan T,

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Analytical chemistry 2016 Oct 4()

Abstract

Photobleaching is a major obstacle in the real-time imaging of biological events, particularly at the single-molecule/particle level. Here, we report a strategy to delay photobleaching of a light-switch complex, [Ru(phen)2dppx]2+, by insertion of a six-cysteine peptide into virus particles. The six-cysteine peptide was inserted into viral protein R of HIV-1, and assembled into infectious HIV-1 viral particles, where it effectively delayed the photobleaching of the [Ru(phen)2dppx]2+ complex used to label viral genomic RNAs. This delay in photobleaching allowed for a mono-fluorescent assay to be constructed for the real-time monitoring of viral uncoating, a poorly understood process. This novel strategy to delay photobleaching in infectious viral particles provides a powerful method to analyze viral uncoating at the single-particle level in real time.

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