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A rhodium-cyanine fluorescent probe: detection and signaling of mismatches in DNA

Nano, Adela and Boynton, Adam N. and Barton, Jacqueline K. (2017) A rhodium-cyanine fluorescent probe: detection and signaling of mismatches in DNA. Journal of the American Chemical Society, 139 (48). pp. 17301-17304. ISSN 0002-7863. PMCID PMC5892186. doi:10.1021/jacs.7b10639. https://resolver.caltech.edu/CaltechAUTHORS:20171115-104208193

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Abstract

We report a bifunctional fluorescent probe that combines a rhodium metalloinsertor with a cyanine dye as the fluorescent reporter. The conjugate shows weak luminescence when free in solution or with well matched DNA but exhibits a significant luminescence increase in the presence of a 27-mer DNA duplex containing a central CC mismatch. DNA photocleavage experiments demonstrate that, upon photoactivation, the conjugate cleaves the DNA backbone specifically near the mismatch site on a 27-mer fragment, consistent with mismatch targeting. Fluorescence titrations with the 27-mer duplex containing the CC mismatch reveal a DNA binding affinity of 3.1 × 10^6 M^(–1), similar to that of other rhodium metalloinsertors. Fluorescence titrations using genomic DNA extracted from various cell lines demonstrate a clear discrimination in fluorescence between those cell lines that are proficient or deficient in mismatch repair. This differential luminescence reflects the sensitive detection of the mismatchrepair-deficient phenotype.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/jacs.7b10639DOIArticle
http://pubs.acs.org/doi/10.1021/jacs.7b10639PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/jacs.7b10639PublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5892186PubMed CentralArticle
ORCID:
AuthorORCID
Nano, Adela0000-0002-1984-5770
Boynton, Adam N.0000-0003-4427-513X
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2017 American Chemical Society. Received: October 5, 2017; Published: November 14, 2017. We are grateful to the NIH (GM33309) for funding this work. We also thank the Moore Foundation, the Beckman Institute Laser Resource Center facilities, and Dr. Jay R. Winkler for assistance. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
NIHGM33309
Issue or Number:48
PubMed Central ID:PMC5892186
DOI:10.1021/jacs.7b10639
Record Number:CaltechAUTHORS:20171115-104208193
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171115-104208193
Official Citation:A Rhodium-Cyanine Fluorescent Probe: Detection and Signaling of Mismatches in DNA. Adela Nano, Adam N. Boynton, and Jacqueline K. Barton. Journal of the American Chemical Society 2017 139 (48), 17301-17304. DOI: 10.1021/jacs.7b10639
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83223
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Nov 2017 21:57
Last Modified:15 Nov 2021 19:56

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