Nucleic acid beacons for long-term real-time intracellular monitoring.
Real time intracellular monitoring of biological molecules inside living cells is important in many biomedical studies and reveals valuable information unobtainable by conventional molecular biology techniques. A variety of methods and molecular probes have been developed, but long term (from a few hours to days) intracellular monitoring with high sensitivity and selectivity is impossible and has not been accomplished. We have used locked nucleic acids (LNA) to engineer novel molecular beacons (MBs) for long-term intracellular monitoring. The LNA-MBs were made of a mixed LNA and DNA bases to have extremely high biostability. The new beacons were tested with MDA-MB-231 cancer cells and used effectively to monitor mRNA expression levels in real-time for 5-24 h. After 24 h inside living cells, the LNA-MBs were still functional, demonstrating a greatly enhanced stability enabling the measurement of intracellular gene expression over an extended period of time.
Wu
Yanrong
Y
Center for Research at Bio/Nano Interface, Department of Chemistry, University of Florida, Gainesville, Florida 32611-7200, USA.
Yang
Chaoyong James
CJ
Moroz
Leonid L
LL
Tan
Weihong
W
D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000737:chemistry; D004355:Drug Stability; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D015336:Molecular Probe Techniques; D009696:Nucleic Acids / Q000737:chemistry* / Q000493:pharmacokinetics; D012333:RNA, Messenger / Q000096:biosynthesis; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics
D018076:DNA, Complementary; D009696:Nucleic Acids; D012333:RNA, Messenger; D013482:Superoxide Dismutase