Volume 61, Issue 2 e202111687
Research Article

Transport of Molecular Cargo by Interaction with Virus-Like Particle RNA

Dr. Soumen Das

Dr. Soumen Das

School of Chemistry and Biochemistry, School of Biological Sciences, Georgia Institute of Technology, 901 Atlantic Dr., Atlanta, GA, 30306 USA

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Dr. Mei-Kwan Yau

Dr. Mei-Kwan Yau

School of Chemistry and Biochemistry, School of Biological Sciences, Georgia Institute of Technology, 901 Atlantic Dr., Atlanta, GA, 30306 USA

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Jeffery Noble

Jeffery Noble

School of Chemistry and Biochemistry, School of Biological Sciences, Georgia Institute of Technology, 901 Atlantic Dr., Atlanta, GA, 30306 USA

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Lucrezia De Pascalis

Lucrezia De Pascalis

School of Chemistry and Biochemistry, School of Biological Sciences, Georgia Institute of Technology, 901 Atlantic Dr., Atlanta, GA, 30306 USA

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Dr. M. G. Finn

Corresponding Author

Dr. M. G. Finn

School of Chemistry and Biochemistry, School of Biological Sciences, Georgia Institute of Technology, 901 Atlantic Dr., Atlanta, GA, 30306 USA

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First published: 30 October 2021
Citations: 8

Graphical Abstract

The binding of intercalating dyes such as thiazole orange (TO) to polynucleotides allows for a simple method to load RNA-rich virus-like particles with molecular cargo, and then to release that cargo by diffusion. TO-labeled molecules come out much slower than they go in because of the high RNA concentration inside the shell; the surprise is how much cargo these easily produced nanocontainers can carry.

Abstract

Virus-like particles (VLPs) derived from Leviviridae virions contain substantial amounts of cellular and plasmid-derived RNA. This encapsidated polynucleotide serves as a reservoir for the efficient binding of the intercalating dye thiazole orange (TO). Polyethylene glycol (PEG) molecules and oligopeptides of varying length, end-functionalized with TO, were loaded into VLPs up to approximately 50 % of the mass of the capsid protein (hundreds to thousands of cargo molecules per particle, depending on size). The kinetics of TO–PEG binding included a significant entropic cost for the reptation of long chains through the capsid pores. Cargo molecules were released over periods of 20–120 hours following simple reversible first-order kinetics in most cases. These observations define a simple general method for the noncovalent packaging, and subsequent release, of functional molecules inside nucleoprotein nanocages in a manner independent of modifications to the capsid protein.

Conflict of interest

The authors declare no conflict of interest.

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