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- Title
A Minimal Load‐and‐Lock Ru<sup>II</sup> Luminescent DNA Probe.
- Authors
Newton, Matthew D.; Fairbanks, Simon D.; Thomas, Jim A.; Rueda, David S.
- Abstract
Threading intercalators bind DNA with high affinities. Here, we describe single‐molecule studies on a cell‐permeant luminescent dinuclear ruthenium(II) complex that has been previously shown to thread only into short, unstable duplex structures. Using optical tweezers and confocal microscopy, we show that this complex threads and locks into force‐extended duplex DNA in a two‐step mechanism. Detailed kinetic studies reveal that an individual stereoisomer of the complex exhibits the highest binding affinity reported for such a mono‐intercalator. This stereoisomer better preserves the biophysical properties of DNA than the widely used SYTOX Orange. Interestingly, threading into torsionally constrained DNA decreases dramatically, but is rescued on negatively supercoiled DNA. Given the "light‐switch" properties of this complex on binding DNA, it can be readily used as a long‐lived luminescent label for duplex or negatively supercoiled DNA through a unique "load‐and‐lock" protocol.
- Subjects
ORANGE SA; LUMINESCENT probes; DNA probes; OPTICAL tweezers; CONFOCAL microscopy; SINGLE molecules
- Publication
Angewandte Chemie, 2021, Vol 133, Issue 38, p21120
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.202108077