Nucleic acid beacons for long-term real-time intracellular monitoring
Nucleic acid beacons for long-term real-time intracellular monitoring
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DOI：
10.1021/ac702637w
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发表时间：
2008-04-15
影响因子：
7.4
通讯作者：
Tan, Weihong
中科院分区：
文献类型：
--
作者：
Wu, Yanrong;Yang, Chaoyong James;Tan, Weihong
活细胞内生物分子的实时监测在许多生物医学研究中非常重要，它揭示了传统分子生物学技术无法获得的有价值的信息。各种方法和分子探针已经开发出来，但长期（从几小时到几天）高灵敏度和选择性的细胞内监测是不可能的，也没有实现。我们已经使用锁定核酸（LNA）来设计用于长期细胞内监测的新型分子信标（mb）。lnamb由LNA和DNA碱基混合制成，具有极高的生物稳定性。新的信标在MDA-MB-231癌细胞中进行了测试，并有效地用于实时监测mRNA表达水平5-24小时。在活细胞内24小时后，lna - mb仍然具有功能，显示出大大增强的稳定性，能够在较长时间内测量细胞内基因表达。
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.