Project Summary
During this project, Electronic BioSicences (EBS) and Professor Meni Wanunu (at Northeastern University) will
develop a true single-molecule, direct, long-lived, RNA sequencer. The developed RNA sequencer will be
capable of sequencing RNA with high-accuracy (>99%), as well as iteratively sequencing various samples and
quantitatively profiling the RNA content of a given sample, as the first technology of its kind. In order to enable
the proposed feat, during this project, we will develop a novel sequencing platform and use it in combination with
newly developed custom enzymes, biological nanopore readers, and sequencing methodology. The
platform/system will feature vastly increased stability and longevity due to the novel platform developments made
during this project. This platform will then be integrated with already developed components, including the
NanopatchTM system sold by EBS, which allows for current monitoring of individual biological nanopores with 10-
fold lower noise than other commercially available instruments, as well as automated methods for planar lipid
bilayer formation and the maintenance of stable protein nanopore insertions, temperature control, and field
programable gate array (FPGA) detection logic to automate decisions on the applied voltage bias. Throughout
this program, we will further optimize the signal-to-noise ratio and temporal resolution of the measurement, and
ultimately the associated based calling. Following the development of the system, we will then demonstrate the
associated sequencing methodology through the sequencing of various RNAs, starting with well-behaved control
sequences before moving to more complicated, biologically relevant RNA sequences. The different RNA
molecules will be sequenced both alone and in mixtures, and the sequence accuracy as well as capture
frequency will be analyzed. We will also determine the ability of the system to handle consecutively loaded
samples. Upon the completing this project, we will have developed, optimized, and fully demonstrated the world’s
first high-accuracy, true single-molecule, direct, long-lived, RNA sequencer. Such technology will transform RNA
sequencing and transcriptomics research by enabling direct, highly accurate, fast, and affordable
characterization of RNA molecules.