Project Summary During this Phase I IMAT SBIR program, Electronic BioSciences, Inc. (EBS), specialists in nanopore platform/methodology developments and high-performance electronics integration, will demonstrate RNA adenosine methylation sequencing by directly differentiating the 3 most abundant adenosine methylations: N6- methyladenosine (m6A), N6,2’-O-dimethyladenosine (m6Am), and N1-methyladenosine (m1A). Current sequencing methods cannot differentiate these three modifications because they are structurally similar, which makes them difficult to differentiate via direct sequencing methods, and because the modification is lost during sequencing-by-synthesis methods that require PCR or amplification. Hence, limitations in current sequencing technology have prevented our ability to assess adenosine methylation as it relates to cancer physiology. By laying the foundation for the development of a sequencing technology capable of direct adenosine methylation detection/differentiation, as will be carried out during this program, we will be able to significantly improve our understanding of how these RNA modification change/progress in cancer and take advantage of this knowledge for new and improved clinical and diagnostic purposes.