The Medusa™ Sequencer: A Sequencing Machine the Size of a Molecule that Could Sequence RNA in a Living Cell
Description of Invention:
Available for licensing and commercial development is the Medusa™ Sequencer, a single-molecule sequencing device that consists of a DNA (or RNA) polymerase attached to a set of four flexible arms. The tip of each arm carries a nonhydrolyzable nucleotide and a spectrally distinct Forster Resonance Energy Transfer (FRET) acceptor fluorophore. A donor fluorophore attached to the polymerase can excite the acceptor fluorophores by FRET. A Medusa™ Sequencer binds to a DNA primer hybridized to the DNA or RNA to be sequenced. The four arms with nucleotide tips "test" the polymerase pocket and the arm that has the nucleotide tip complementary to the unknown base of the sequence will dwell longer than the other three that are not complementary. However, the polymerase will not incorporate the nucleotide on the tip of the arm into the nascent strand because the nucleotide is nonhydrolyzable. FRET between the donor and the acceptor fluorophore at the arm tip produces a characteristic spectrum that identifies the bound base. Free hydrolyzable dNTPs (or NTPs) allow the Medusa™ Sequencer to step forward. The series of FRET signals reveals the unknown nucleotide sequence. A Medusa™ Sequencer could also be injected into a cell to read mRNA sequences inside a living organism. Coded versions of the Medusa™ Sequencer can signal when the device has been damaged.
The benefits of the Medusa™ Sequencer include: a) simplicity, only one reagent required; b) accuracy for counting individual mRNAs or DNAs; c) low error rate per base, and this can be improved by modifying the polymerase; d) speed, a single microscope can be used to obtain many sequences in parallel; e) exceptionally low cost per sequencing device; and d) could be used in the clinic along with sequence walkers to analyze patient's genetic diseases (e.g. Medical Applications of Sequence Walkers: ABCR Mutation G863A, http://www.ccrnp.ncifcrf.gov/~toms/g863a.html).
Inventors:
Thomas D. Schneider (NCI) Ilya G. Lyakhov (NCI) Danielle Needle (NCI)
Patent Status:
DHHS Reference No. E-194-2005/0 --
U.S. Provisional Application No. 60/749,729 filed 12 Dec 2005
PCT Application No. PCT/US2006/047534 filed 12 Dec 2006, which published as WO 2007/070572 on 21 Jun 2007
U.S. Patent Application No. 12/095,973 filed 03 Jun 2008
Related Technologies: DHHS Reference No. E-195-2005/0 --
U.S. Patent Application No. 11/638,160 filed 12 Dec 2006, claiming priority to 12 Dec 2005
Licensing Status: Available for non-exclusive or exclusive licensing.
Collaborative Research Opportunity:
The National Institutes of Health, National Cancer Institute, Center for Cancer Research Nanobiology Program is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize the Medusa™ Sequencer. Please contact Melissa Maderia at 301/846-5465 (phone), 301/846-6820 (fax), maderiam@mail.nih.gov (e-mail) for more information.
Portfolios: Miscellaneous Internal Medicine Infectious Diseases Gene Based Therapies Devices/Instrumentation Cancer
Cancer -Diagnostics-In Vitro-DNA Based Cancer -Diagnostics-In Vivo-Conjugate Chemistry Cancer -Diagnostics Devices/Instrumentation-Diagnostics Gene Based Therapies -Research Materials Infectious Diseases -Diagnostics Internal Medicine-Diagnostics Internal Medicine-Research Materials Miscellaneous-Miscellaneous
For Additional Information Please Contact: Cristina Thalhammer-Reyero PhD MBA
NIH Office of Technology Transfer
6011 Executive Blvd, Suite 325
Rockville, MD 20852-3804
Phone: (301) 435-4507
Email: thalhamc@mail.nih.gov
Fax: (301) 402-0220
