ArticleRNA (New York, N.Y.)2010
Monitoring viral RNA in infected cells with LNA flow-FISH.
Article in RNA (New York, N.Y.), 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Shedding Light on Intracellular Proteins using Flow Cytometry.Cell biochemistry and biophysics · 2024Review
- Flow-FISH as a Tool for Studying Bacteria, Fungi and Viruses.Biotech (Basel (Switzerland)) · 2021Review
- Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies.Virology · 2016Article
- Flow Cytometric Characterization of Antigen-Specific T Cells Based on RNA and Its Advantages in Detecting Infections and Immunological Disorders.Critical reviews in immunology · 2016Article
- Marine viruses, a genetic reservoir revealed by targeted viromics.The ISME journal · 2014Article
- Transcriptional profiling of cells sorted by RNA abundance.Nature methods · 2014Article
- Assessment of different permeabilization methods of minimizing damage to the adherent cells for detection of intracellular RNA by flow cytometry.Avicenna journal of medical biotechnology · 2014Article
- Article
- Detection of single influenza viral RNA in cells using a polymeric sequence probe.Analytical chemistry · 2012Article
- Tracking expression and subcellular localization of RNA and protein species using high-throughput single cell imaging flow cytometry.RNA (New York, N.Y.) · 2012Article
- Locked nucleic acid flow cytometry-fluorescence in situ hybridization (LNA flow-FISH): a method for bacterial small RNA detection.Journal of visualized experiments : JoVE · 2012Article
- Locked nucleic acid and flow cytometry-fluorescence in situ hybridization for the detection of bacterial small noncoding RNAs.Applied and environmental microbiology · 2012Article
- Co-expression vs. co-infection using baculovirus expression vectors in insect cell culture: Benefits and drawbacks.Biotechnology advancesReview
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We previously showed the feasibility of using locked nucleic acid (LNA) for flow cytometric-fluorescence in situ hybridization (LNA flow-FISH) detection of a target cellular mRNA. Here we demonstrate how the method can be used to monitor viral RNA in infected cells. We compared the results of the LNA flow-FISH with other methods of quantifying virus replication, including the use of an enhanced green fluorescent protein (EGFP) viral construct and quantitative reverse-transcription polymerase chain reaction. We found that an LNA probe complementary to Sindbis virus RNA is able to track the increase in viral RNA over time in early infection. In addition, this method is comparable to the EGFP construct in sensitivity, with both peaking around 3 h and at the same level of infected cells. Finally, we observed that the LNA flow-FISH method responds to the decrease in levels of viral RNA caused by antiviral medication. This technique represents a straightforward way to monitor viral infection in cells and is easily applicable to any virus.
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