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Simultaneous Protein and RNA Analysis in Single Extracellular Vesicles, Including Viruses
Zach Troyer1, Olesia Gololobova1,2, Aakash Koppula3
1Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, United States.
ACS Nano
|September 22, 2024
Summary
A new method called SPIRFISH can now distinguish infectious human immunodeficiency virus (HIV) from extracellular vesicles (EVs) by detecting both viral protein and genomic RNA in single particles. This breakthrough aids in understanding viral dynamics and advancing EV therapeutics.
Area of Science:
- Virology
- Nanotechnology
- Biochemistry
Background:
- Distinguishing infectious human immunodeficiency virus (HIV) from extracellular vesicles (EVs) is challenging due to similar physical properties.
- Current single-particle analysis methods primarily detect proteins, failing to differentiate between infectious virions, nonviral EVs, and hybrid EVs.
Purpose of the Study:
- To develop a high-throughput method for simultaneous detection of viral protein and genomic RNA in single nanoparticles.
- To enable accurate resolution of infectious HIV virions from EVs and noninfectious particles.
Main Methods:
- Introduction of SPIRFISH (Single Particle Interferometric Reflectance Imaging Sensor with Single-molecule Fluorescence In Situ Hybridization).
- Simultaneous detection of HIV-1 envelope protein gp120 and genomic RNA within individual virions and EVs.
Main Results:
- SPIRFISH successfully resolved infectious HIV-1 virions from EVs and noninfectious virions by detecting both protein and genomic RNA.
- The method demonstrated capability in detecting specific RNAs within EVs, highlighting potential for EV therapeutics.
Conclusions:
- SPIRFISH provides a novel high-throughput approach for single-particle protein and RNA analysis.
- This technique has significant implications for studying viral infections, differentiating viral particles from EVs, and advancing RNA-based EV therapeutics.
