Selective Detection of 8-Oxo-2′-deoxyguanosine in Single-Stranded DNA via Nanopore Sensing Approach

Abstract

We have developed a nanopore sensing approach for the selective detection of 8-oxo-2'-deoxyguanosine (8-oxoG) in single-stranded DNA. First, 1,12-dodecanediamine is coupled with 8-oxoG-containing DNA molecules in high yield which leaves a free amine group for subsequent attaching of an adamantane moiety. After incubation with cucurbit[7]uril, the host-guest complex-modified DNA hybrid is translocated through an α-hemolysin nanopore. Highly characteristic events can be recorded and used to quantify the 8-oxoG-DNA content in a DNA mixture. Compared with the existing methods, this study provides a reliable, quick, and low-cost approach for the detection of 8-oxoG site in single-stranded DNA at the single-molecule level, particularly suitable for high-throughput screening of a massive number of samples.

Quantification and mapping of DNA modifications

“…Translocation of this DNA hybrid generates current signatures reflective of the presence of OG in DNA. 224 …”

Cucurbiturils in nucleic acids research

“…Since 2015, H.-C. Wu and co-workers are developing a versatile method for nanopore sensing of biologically related analytes based on DNA-CB [7] host-guest probes. [75][76][77][78][79][80][81][82][83] In the method developed by H.-C. Wu and co-workers, introduction of the analyte leads to the release or in situ formation of the single-stranded DNA (ssDNA) bearing a ferrocene-CB [7] or adamantane-CB [7] complex, referred to as a DNA probe. Threading of the DNA probe through the a-hemolysin (aHL) nanopore results in the dissociation of the host-guest complex and liberation of CB [7] (Fig.…”

Measuring Enzymatic Activities with Nanopores

“…Our group have been using DNA probes for nanopore sensing for more than one decade. By exploiting the current signal changes caused by DNA conformational changes or DNA modifications during their translocation through protein nanopores, we have developed a series of approaches for sensing a wide variety of different analytes . To measure enzymatic activities, we attached the peptide substrate to a DNA molecule to form a hybrid probe.…”

“…By exploiting the current signal changes caused by DNA conformational changes or DNA modifications during their translocation through protein nanopores, we have developed a series of approaches for sensing a wide variety of different analytes. [45][46][47][48][49][50][51][52] To measure enzymatic activities, we attached the peptide substrate to a DNA molecule to form a hybrid probe. We managed to amplify the current difference before and after enzymatic cleavage by the introduction of cucurbit [7]uril (CB [7]) which is known to have strong affinities with peptides containing N-terminal phenylalanine ( Figure 5A).…”