A lentiviral vector encoding two fluorescent proteins enables imaging of adenoviral infection via adenovirus-encoded miRNAs in single living cells.
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.
Kato
Yoshio
Y
Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Japan. y-kato@aist.go.jp
Sawata
Shinya Y
SY
Inoue
Atsushi
A
D000256:Adenoviridae / Q000235:genetics* / Q000302:isolation & purification*; D000257:Adenoviridae Infections / Q000821:virology*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics
D008164:Luminescent Proteins; D035683:MicroRNAs