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Review
. 2020 Aug 21;12(9):917.
doi: 10.3390/v12090917.

Purification Methods and the Presence of RNA in Virus Particles and Extracellular Vesicles

Affiliations

Affiliations

  • 1 Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.
  • 2 Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.
Review

Purification Methods and the Presence of RNA in Virus Particles and Extracellular Vesicles

Yijun Zhou et al. Viruses. .
. 2020 Aug 21;12(9):917.
doi: 10.3390/v12090917.

Affiliations

  • 1 Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.
  • 2 Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.

Abstract

The fields of extracellular vesicles (EV) and virus infections are marred in a debate on whether a particular mRNA or non-coding RNA (i.e., miRNA) is packaged into a virus particle or copurifying EV and similarly, whether a particular mRNA or non-coding RNA is contained in meaningful numbers within an EV. Key in settling this debate, is whether the purification methods are adequate to separate virus particles, EV and contaminant soluble RNA and RNA:protein complexes. Differential centrifugation/ultracentrifugation and precipitating agents like polyethylene glycol are widely utilized for both EV and virus purifications. EV are known to co-sediment with virions and other particulates, such as defective interfering particles and protein aggregates. Here, we discuss how encased RNAs from a heterogeneous mixture of particles can be distinguished by different purification methods. This is particularly important for subsequent interpretation of whether the RNA associated phenotype is contributed solely by virus or EV particles or a mixture of both. We also discuss the discrepancy of miRNA abundance in EV from different input material.

Keywords: exosomes; extracellular vesicles; herpesviruses; virion RNA.

PubMed Disclaimer

Conflict of interest statement

There are no conflicts of interest. D.P.D receives kind contributions from GE Health Science and R.P.M and D.P.D have developed tangible intellectual property in the field.

Figures

Figure 1

Figure 1

Extracellular vesicles (EV) and virus…

Figure 1

Extracellular vesicles (EV) and virus particles share similar vesicular budding process and composition,…

Figure 1
Extracellular vesicles (EV) and virus particles share similar vesicular budding process and composition, including proteins, nucleic acids, and lipids. (A) Microvesicles bud off at the plasma membrane, similar to the canonical human immunodeficiency viruses (HIV) budding process. Exosomes originate from inward budding of the late endosome into multivesicular body (MVB) and later release at the plasma membrane, similar to the canonical enveloped hepatitis A viruses (HAV) budding process. (B) Extracellular vesicles may carry makers like tetraspanins, esterases, Alix, and Tsg101 [1]. The encased nucleic acids are protected from nucleases. (C). A virus particle consists of an envelope, capsid, tegument, and viral genome.
Figure 2

Figure 2

70% of all miRNAs in…

Figure 2

70% of all miRNAs in infected human vein endothelial cells (HUVECs) come from…

Figure 2
70% of all miRNAs in infected human vein endothelial cells (HUVECs) come from KSHV.

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