Direct Sequencing of 5-Methylcytosine and 5-Hydroxymethylcytosine at Single-Base Resolution Unravels Their Distinct Roles in Alzheimer's Disease.

ABSTRACT: Alzheimer's disease (AD) is associated with genetic risk factors and widespread epigenetic alterations. 5-Hydroxymethylcytosine (5hmC), an oxidized derivative of 5-methylcytosine (5mC), constitutes up to 20% of 5mC in neuronal DNA and is implicated in aging and neurodegeneration. However, the precise roles of DNA modifications in AD remain unclear, partly due to the lack of accurate detection methods. Here, two orthogonal sequencing methods are introduced: CMD1-Deaminase sequencing (CD-seq) and CMD1-TET bisulfite sequencing (CT-seq), which enable direct, independent detection of 5mC. When combined with APOBEC-coupled epigenetic sequencing (ACE-seq) or TET-assisted bisulfite sequencing (TAB-seq) for 5hmC mapping, these techniques provide base-resolution, subtraction-free profiling of DNA mo

SUBMITTER: Wang ZX 

PROVIDER: S-EPMC12520568 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature