A Cleavable Molecular Beacon for Hg2+ Detection based on Phosphorothioate RNA Modifications
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Mercury is a highly toxic heavy metal, and detection of Hg2+ by biosensors has attracted extensive research interest in the past decade. In particular, a number of DNA-based sensing strategies have been developed. Well-known examples include thymine-Hg2+ interactions and Hg2+-activated DNAzymes. However, these mechanisms are highly dependent on buffer conditions or require hybridization with another DNA strand. Herein, we report a new mechanism based on Hg2+-induced cleavage of phosphorothioate (PS) modified RNA. Among the various metal ions tested, Hg2+ induced the most significant cleavage (∼16%), while other metals cleaved less than 2% of the same substrate. The uncleaved substrate undergoes desulfurization in the presence of Hg2+. This cleavage reaction yields a similar amount of product from pH 3.5 to 7 and in the temperature range between 20 and 90 °C. Various PS RNA junctions can be cleaved with a similar efficiency, but PS DNA junctions cannot be cleaved. A molecular beacon containing three PS RNA modifications is designed, detecting Hg2+ down to 1.7 nM with excellent selectivity. This sensor can also detect Hg2+ in the Lake Ontario water sample, although its response is significantly masked by fish tissues.
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Po-Jung Jimmy Huang, Feng Wang, Juewen Liu (2015). A Cleavable Molecular Beacon for Hg2+ Detection based on Phosphorothioate RNA Modifications. UWSpace. http://hdl.handle.net/10012/11345