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Long-lived Platform Development for Exonuclease-Based Sequencing

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R41HG012163-01
Agency Tracking Number: R41HG012163
Amount: $399,999.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: 172
Solicitation Number: PA20-265
Solicitation Year: 2020
Award Year: 2021
Award Start Date (Proposal Award Date): 2021-09-01
Award End Date (Contract End Date): 2022-08-31
Small Business Information
San Diego, CA 92121-4206
United States
DUNS: 129852864
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 (801) 582-0146
Business Contact
Phone: (858) 412-1800
Research Institution
360 HUNTINGTON AVE, 177-500
BOSTON, MA 02115-5005
United States

 Nonprofit College or University

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 (andgt;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.Project Narrative
The technology developed during this project will be the world’s first, high-accuracy, true single-molecule, direct,
long-lived, RNA sequencer, suitable for profiling RNA samples and identifying the associated critical
modifications and biomarkers. Through this development a new and detailed understanding of RNA and its
regulation will be possible. A technology capable of directly sequencing RNA with high accuracy, a feat no other
technology can perform, will transform the understanding of the transcriptome and enable new diagnostics,
prognostics, and therapeutic interventions, ultimately improving public health.

* Information listed above is at the time of submission. *

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