Hybridization-Sensitive Fluorescent Oligonucleotide Probe Conjugated with a Bulky Module for Compartment-Specific mRNA Monitoring in a Living Cell
mediaposted on 2015-03-18, 00:00 authored by Gosuke Hayashi, Masafumi Yanase, Katsuya Takeda, Daisuke Sakakibara, Ryosuke Sakamoto, Dan Ohtan Wang, Akimitsu Okamoto
Live-cell RNA imaging at specific intracellular locations is technically limited because of the diffusive nature of small oligonucleotide probes. The bulky fluorescent light-up probes that possess streptavidin or gold nanoparticles at the end of oligonucleotides were designed and synthesized. The bulky probes allowed nucleus- and cytoplasm-selective monitoring of endogenous mRNAs through nuclear and cytoplasmic microinjection, respectively. Simultaneous use of bulky and unbulky probes conjugated with different fluorescent dyes enabled dual color imaging of mRNAs present in nucleus and cytoplasm. Furthermore, we observed that the fluorescence near the cell edge in a living HeLa cell traveled over time in coordination with the dynamic formation and deformation of the pseudopodial protrusions after lipofection of the bulky probes.