Rhodium-cyanine fluorescent probes for detection and signaling of mismatches in DNA
- Creators
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Nano, Adela
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Barton, Jacqueline
Abstract
Mismatched (non-Watson-Crick) base pair damage in DNA occurs naturally from errors during the replication process. Deficiencies in the mismatch repair (MMR) machinery, a DNA repair pathway, strongly predispose cells to cancer development. Therefore, efficiently detecting DNA mismatches will greatly enable early detection of MMR-deficient precancerous cells. Herein, we report the synthesis and characterization of a bifunctional fluorescent probe that combines a rhodium metalloinsertor with indol trimethine cyanine, Cy(3), the luminescent reporter, via a PEG-type linker. The conjugate displays low luminescence when free in soln. or in the presence of well-matched DNA but exhibits a luminescence increase up to 9-fold in the presence of a 27-mer oligonucleotide contg. a central CC mismatch. DNA photocleavage expts. demonstrate that upon photoactivation, the conjugate can cleave the DNA backbone near the mismatch site on a 27-mer oligonucleotide, thus providing further evidence for mismatch targeting. Fluorescence titrns. of the Rh conjugate with genomic DNA (gDNA) extd. from MMR-deficient and MMR-proficient HCT116 cell lines show a luminescence differential between gDNA from MMR-deficient and - proficient cell lines, reflecting the sensitive detection of differences in mismatch frequency.
Additional Information
© 2017 American Chemical Society.Additional details
- Eprint ID
- 81350
- Resolver ID
- CaltechAUTHORS:20170912-100733809
- Created
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2017-09-12Created from EPrint's datestamp field
- Updated
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2019-10-03Created from EPrint's last_modified field