Project Summary
Nanopore-based technology has the potential to be the future of comprehensive RNA sequencing, i.e.,
transcriptomics and epitranscriptomics, as it is the only viable path to directly characterizing and profiling long
RNA strands without reverse transcription or processive enzymes that fail to traverse certain sequences. The
direct characterization enabled by nanopore sequencing will also ultimately lead to detection and
characterization of modified bases, a critical requirement for epitranscriptomics. The presently available, state-
of-the art RNA sequencing technologies are limited by the method, with the input requiring cDNA intermediates
and/or large amounts of costly sample, and the readout featuring a compromise between short read length and
low-accuracy. During this program, Electronic BioSciences (EBS) aims to develop a completely new long-read,
extremely high-accuracy nanopore-based sequencing technology that will enable de novo epitranscriptomic
sequencing, capable of inherent sequence cross-validation and expanded sensitivity for more accurate and
versatile RNA sequencing. During this Phase I project, we will build and multiplex an entirely new system, fully
assess and optimize the associated workflow/methodology to sequence canonical nucleotides and specific
clinically relevant RNA modifications, and demonstrate initial sequencing. At the conclusion of this project, we
will have successfully demonstrated and developed an entirely new sequencing system prototype that
overcomes the known challenges limiting current RNA sequencing technologies (including other nanopore-
based sequencing approaches). The resulting technology development will be a label-free, long-read (>kbases),
electronic readout method, capable of providing a new and detailed understanding of RNA and its regulation.