Sequencing of N6-methyl-deoxyadenosine at single-base resolution across the mammalian genome.
Mol Cell
; 84(3): 596-610.e6, 2024 Feb 01.
Article
in En
| MEDLINE
| ID: mdl-38215754
ABSTRACT
Although DNA N6-methyl-deoxyadenosine (6mA) is abundant in bacteria and protists, its presence and function in mammalian genomes have been less clear. We present Direct-Read 6mA sequencing (DR-6mA-seq), an antibody-independent method, to measure 6mA at base resolution. DR-6mA-seq employs a unique mutation-based strategy to reveal 6mA sites as misincorporation signatures without any chemical or enzymatic modulation of 6mA. We validated DR-6mA-seq through the successful mapping of the well-characterized G(6mA)TC motif in the E. coli DNA. As expected, when applying DR-6mA-seq to mammalian systems, we found that genomic DNA (gDNA) 6mA abundance is generally low in most mammalian tissues and cells; however, we did observe distinct gDNA 6mA sites in mouse testis and glioblastoma cells. DR-6mA-seq provides an enabling tool to detect 6mA at single-base resolution for a comprehensive understanding of DNA 6mA in eukaryotes.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
DNA Methylation
/
Escherichia coli
Limits:
Animals
Language:
En
Journal:
Mol Cell
/
Mol. cell
/
Molecular cell
Journal subject:
BIOLOGIA MOLECULAR
Year:
2024
Type:
Article
Affiliation country:
United States