Detection of 5-methylcytosine and 5-hydroxymethylcytosine in DNA via host-guest interactions inside α-hemolysin nanopores

Tao Zeng, Lei Liu, Ting Li, Yuru Li, Juan Gao, Yuliang Zhao, Hai Chen Wu

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Cytosine methylation and hydroxymethylation are both important epigenetic modifications of DNA in mammalian cells. Therefore, profiling DNA (hydroxy)methylation across the genome is vital for understanding their roles in gene regulation. Here, we report a nanopore-based approach for quick and reliable detection of 5-methylcytosine and 5-hydroxymethylcytosine in DNA at the single-molecule level. The single-stranded DNA containing 5-methylcytosine or 5-hydroxymethylcytosine was first selectively modified on the epigenetic base to attach a host-guest complex. Threading of the modified DNA molecules through α-hemolysin nanopores causes unbinding of the host-guest complex and generates highly characteristic current signatures. Statistical analysis of the signature events affords quantitative information about 5-methylcytosine and 5-hydroxymethylcytosine in DNA. Our results suggest that other DNA modifications could also be detected with the developed method. Furthermore, we anticipate our nanopore sensing strategy to be generally useful in biochemical analysis and to find applications in the early diagnosis of diseases.

Original languageEnglish (US)
Pages (from-to)5628-5634
Number of pages7
JournalChemical Science
Volume6
Issue number10
DOIs
StatePublished - Jun 24 2015

ASJC Scopus subject areas

  • Chemistry(all)

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