From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging
Abstract
:1. Introduction
2. In Vitro Gel-Based Approaches: Northern Blot and Ribonuclease Protection Assay
2.1. The Northern Blot
2.2. The RPA
2.2.1. Sensitivity
2.2.2. Specificity and Accuracy
2.2.3. Multiplexing
2.3. Applications of Northern Blot and RPA
3. Quantitative RT-PCR
3.1. Absolute versus Relative Quantification
3.2. Choice of Fluorescence Approach
3.3. Priming Strategy
3.4. Quality of Templates
3.5. RT-qPCR to Monitor Retroviral RNA Encapsidation
3.6. Advantages of RT-qPCR.
4. Fluorescence Microscopy of Fixed- or Live-Cells
4.1. FISH
4.1.1. Methodology
4.1.2. Applications of FISH in the Study of Retroviral RNA Encapsidation
4.2. Live-Cell Imaging to Study Viral RNA Packaging
4.3. FISH versus Live-Cell Imaging
5. RNA Sequencing Opens New Perspectives for Studying the Virion Transcriptome
6. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Ferrer, M.; Henriet, S.; Chamontin, C.; Lainé, S.; Mougel, M. From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging. Viruses 2016, 8, 239. https://doi.org/10.3390/v8080239
Ferrer M, Henriet S, Chamontin C, Lainé S, Mougel M. From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging. Viruses. 2016; 8(8):239. https://doi.org/10.3390/v8080239
Chicago/Turabian StyleFerrer, Mireia, Simon Henriet, Célia Chamontin, Sébastien Lainé, and Marylène Mougel. 2016. "From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging" Viruses 8, no. 8: 239. https://doi.org/10.3390/v8080239