Method for nanopore rna characterisation
AssigneeOxford Nanopore Technologies PLC
InventorsDaniel Ryan Garalde, Andrew John Heron, Lakmal Nishantha Jayasinghe, Daniel John Turner, James White
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Abstract
The invention relates to a new method of characterising a target RNA polynucleotide by taking one or more measurements as the target RNA polynucleotide moves with respect to a transmembrane pore. The movement is controlled by a DNA helicase. The invention also relates to a modified RNA construct wherein the RNA polynucleotide has been modified to increase DNA helicase binding thereto.
Claims28
A method of characterising a target RNA polynucleotide comprising:
- a) providing (i) an RNA polynucleotide wherein the RNA polynucleotide is modified to comprise a non-RNA polynucleotide and (ii) a DNA helicase enzyme;
- b) contacting the RNA polynucleotide and DNA helicase enzyme provided in a) with a transmembrane pore such that the DNA helicase controls the movement of the RNA polynucleotide through the transmembrane pore;
- c) taking one or more measurements as the RNA polynucleotide moves with respect to the transmembrane pore, wherein the measurements are indicative of one or more characteristics of the RNA polynucleotide, and thereby characterising the target RNA polynucleotide.
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A method of moving a target RNA polynucleotide with respect to a transmembrane pore when the movement is controlled by a DNA helicase enzyme, comprising:
- a) providing (i) an RNA polynucleotide wherein the RNA polynucleotide is modified to comprise a non-RNA polynucleotide and a (ii) a DNA helicase enzyme;
- b) contacting the RNA polynucleotide and DNA helicase enzyme provided in a) with a transmembrane pore such that the DNA helicase controls the movement of the RNA polynucleotide with respect to the transmembrane pore.
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A polynucleotide comprising an RNA polynucleotide and DNA polynucleotide wherein the DNA polynucleotide comprises or comprises only a DNA helicase binding site, optionally wherein the polynucleotide further comprises a leader sequence.
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A combination of an RNA polynucleotide and a DNA helicase in which a part of the RNA polynucleotide has been modified to comprise a non-RNA polynucleotide and interact with the DNA helicase.
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Dependencies are inferred from the claim text.