A lentiviral vector encoding two fluorescent proteins enables imaging of adenoviral infection via adenovirus-encoded miRNAs in single living cells.

Abstract

MicroRNAs (miRNAs) are non-coding small RNAs that have been found in various kinds of eukaryotes and viruses. Recently, adenovirus non-coding RNAs, VA RNAs, have been reported to generate miRNAs. Here, we developed a lentiviral vector for monitoring adenovirus-derived miRNAs in living cells. By using red and green fluorescent proteins under the control of bi-directional two distinct promoters, adenoviral infection and consequent miRNA expression was successfully visualized and quantified by the reduction in green fluorescence when 3'-untranslated regions were connected to the target sequences of the adenovirus-derived miRNAs. Our functional analysis using a lentiviral vector is a useful method to examine the activity of miRNA in living mammalian cells.

• Authors: Yoshio Kato, Shinya Y Sawata, Atsushi Inoue
• Journal: Journal of biochemistry (J Biochem) Vol. 147 Issue 1 Pg. 63-71 (Jan 2010) ISSN: 1756-2651 [Electronic] England
• PMID: 19762345 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Luminescent Proteins
• MicroRNAs

Topics

• Adenoviridae (genetics, isolation & purification)
• Adenoviridae Infections (virology)
• Cell Line
• Genetic Vectors (genetics)
• Humans
• Luminescent Proteins (analysis, genetics, metabolism)
• MicroRNAs (analysis, genetics)
• Promoter Regions, Genetic (genetics)