Spatially resolved m6A profiling using m6A-ARTR-DBiT

N6-methyladenosine (m6A) on RNA plays diverse regulatory roles, yet its spatial distribution within tissues remains largely unexplored. Here, we develop a spatial m6A profiling assay leveraging reverse transcription-based detection and deterministic barcoding in tissue (m6A-ARTR-DBiT), enabling spatially resolved sequencing of m6A within the native tissue context. Application of m6A-ARTR-DBiT to embryonic and adult mouse brain generated high-resolution whole transcriptome spatial m6A maps, offering insights into region-specific m6A modification. Overall design: We developed an m6A profiling assay leveraging reverse transcription and deterministic barcoding in tissue (m6A-ARTR-DBiT) for spatial exploration of m6A modifications. We benchmarked this method on cell lines, demonstrating its capability to detect m6A modifications, and further applied m6A-ARTR-DBiT to mouse embryonic and brain tissues to generate high-resolution, whole-transcriptome spatial m6A maps.