Open AccessCreation of RNA molecules that recognize the oxidative lesion 7,8-dihydro-8-hydroxy-2′-deoxyguanosine (8-oxodG) in DNA
Author(s) -
Stacia M. Rink,
Jiang-Cheng Shen,
Lawrence A. Loeb
Publication year - 1998
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.95.20.11619
Subject(s) - dna , rna , deoxyguanosine , 8 hydroxy 2' deoxyguanosine , dna oxidation , oligonucleotide , dna damage , microbiology and biotechnology , biochemistry , transcription (linguistics) , nucleotide , chemistry , nucleic acid , biology , gene , linguistics , philosophy
We usedin vitro evolution to obtain RNA molecules that specifically recognize and bind with high affinity to the oxidative lesion 7,8-dihydro-8-hydroxy-2′-deoxyguanosine (8-oxodG) in DNA. A pool of ≈1015 RNA molecules containing a random insert of 45 nucleotides in length was subject to 10 successive rounds of chromatographic enrichment using an 8-oxodG affinity matrix, reverse transcription, PCR amplification, and RNA synthesis. Selected RNA molecules bind to 8-oxodG located at the 3′ terminus (K d ≤ 270 nM) or in the center (K d ≤ 2.8 μM) of a 19-nt strand of DNA, with no detectable affinity for the corresponding dG-containing DNA sequences. These 8-oxodG-binding RNAs will be used to monitor levels of 8-oxodG in DNA from biological sources and should provide a unique method for evaluating oxygen-mediated DNA damage. This approach should be applicable for the creation of RNA molecules that can bind to and identify the different modifications of DNA produced by a variety of environmental agents.