Please use this identifier to cite or link to this item: https://doi.org/10.1093/nar/gkab515
Title: High-affinity five/six-letter DNA aptamers with superior specificity enabling the detection of dengue NS1 protein variants beyond the serotype identification
Authors: Matsunaga, Ken-Ichiro
Kimoto, Michiko
Lim, Vanessa Weixun
Tan, Hui Pen
Wong, Yu Qian
Sun, William 
Vasoo, Shawn
Leo, Yee Sin 
Hirao, Ichiro
Issue Date: 25-Jun-2021
Publisher: Oxford University Press
Citation: Matsunaga, Ken-Ichiro, Kimoto, Michiko, Lim, Vanessa Weixun, Tan, Hui Pen, Wong, Yu Qian, Sun, William, Vasoo, Shawn, Leo, Yee Sin, Hirao, Ichiro (2021-06-25). High-affinity five/six-letter DNA aptamers with superior specificity enabling the detection of dengue NS1 protein variants beyond the serotype identification. Nucleic Acids Research 49 (20) : 11407-11424. ScholarBank@NUS Repository. https://doi.org/10.1093/nar/gkab515
Rights: Attribution 4.0 International
Abstract: Genetic alphabet expansion of DNA by introducing unnatural bases (UBs), as a fifth letter, dramatically augments the affinities of DNA aptamers that bind to target proteins. To determine whether UB-containing DNA (UB-DNA) aptamers obtained by affinity selection could spontaneously achieve high specificity, we have generated a series of UB-DNA aptamers (KD: 27-182 pM) targeting each of four dengue non-structural protein 1 (DEN-NS1) serotypes. The specificity of each aptamer is remarkably high, and the aptamers can recognize the subtle variants of DEN-NS1 with at least 96.9% amino acid sequence identity, beyond the capability of serotype identification (69-80% sequence identities). Our UB-DNA aptamers specifically identified two major variants of dengue serotype 1 with 10-amino acid differences in the DEN-NS1 protein (352 aa) in Singaporeans' clinical samples. These results suggest that the high-affinity UB-DNA aptamers generated by affinity selection also acquire high target specificity. Intriguingly, one of the aptamers contained two different UBs as fifth and sixth letters, which are essential for the tight binding to the target. These two types of unnatural bases with distinct physicochemical properties profoundly expand the potential of DNA aptamers. Detection methods incorporating the UB-DNA aptamers will facilitate precise diagnoses of viral infections and other diseases. © 2021 The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.
Source Title: Nucleic Acids Research
URI: https://scholarbank.nus.edu.sg/handle/10635/232367
ISSN: 0305-1048
DOI: 10.1093/nar/gkab515
Rights: Attribution 4.0 International
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