Nucleic acid beacons for long-term real-time intracellular monitoring

被引:60
作者
Wu, Yanrong
Yang, Chaoyong James
Moroz, Leonid L.
Tan, Weihong [1 ]
机构
[1] Univ Florida, Ctr Res Bio Nano Interface, Dept Chem, Dept Neurosci,UF Genet Inst, Gainesville, FL 32611 USA
[2] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA
[3] Univ Florida, Shands Canc Ctr, Gainesville, FL 32611 USA
关键词
D O I
10.1021/ac702637w
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 LNAMBs 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.
引用
收藏
页码:3025 / 3028
页数:4
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