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  • DiMeLo-seq: a long-read, single-molecule method for mapping protein-DNA interactions genome wide. Nature methods Altemose, N., Maslan, A., Smith, O. K., Sundararajan, K., Brown, R. R., Mishra, R., Detweiler, A. M., Neff, N., Miga, K. H., Straight, A. F., Streets, A. 2022

    Abstract

    Studies of genome regulation routinely use high-throughput DNA sequencing approaches to determine where specific proteins interact with DNA, and they rely on DNA amplification and short-read sequencing, limiting their quantitative application in complex genomic regions. To address these limitations, we developed directed methylation with long-read sequencing (DiMeLo-seq), which uses antibody-tethered enzymes to methylate DNA near a target protein's binding sites in situ. These exogenous methylation marks are then detected simultaneously with endogenous CpG methylation on unamplified DNA using long-read, single-molecule sequencing technologies. We optimized and benchmarked DiMeLo-seq by mapping chromatin-binding proteins and histone modifications across the human genome. Furthermore, we identified where centromere protein A localizes within highly repetitive regions that were unmappable with short sequencing reads, and we estimated the density of centromere protein A molecules along single chromatin fibers. DiMeLo-seq is a versatile method that provides multimodal, genome-wide information for investigating protein-DNA interactions.

    View details for DOI 10.1038/s41592-022-01475-6

    View details for PubMedID 35396487