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US 11,021,747 B2Granted
Method for nanopore RNA characterisation
AssigneeOxford Nanopore Technologies PLC
InventorsDaniel Ryan Garalde, Andrew John Heron, Lakmal 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.
Claims19
A method of characterising a target RNA polynucleotide comprising:
- (a) providing (i) an RNA polynucleotide that is modified to comprise a single stranded DNA polynucleotide and (ii) a helicase enzyme, wherein the single stranded DNA polynucleotide provides a binding site for the helicase enzyme;
- (b) contacting the RNA polynucleotide and helicase enzyme provided in (a) with a transmembrane pore such that the helicase enzyme 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 one or more measurements are indicative of one or more characteristics of the RNA polynucleotide, and
- thereby characterising the target RNA polynucleotide.
1 more element
A method of characterising an RNA polynucleotide comprising:
- (a) providing (i) an RNA polynucleotide that is modified to comprise a single stranded DNA polynucleotide, (ii) a helicase enzyme, and (iii) one or more molecular brakes that are derived from helicases and are modified such that they bind the polynucleotide but do not function as a helicase;
- (b) contacting the modified RNA polynucleotide and helicase enzyme provided in (a) with a transmembrane pore such that the helicase controls the movement of the RNA polynucleotide through the transmembrane pore;
- (c) taking one or more measurements as the modified RNA polynucleotide moves with respect to the transmembrane pore, wherein the one or more measurements are indicative of one or more characteristics of the RNA polynucleotide, and
- thereby characterising the RNA polynucleotide.
1 more element
Dependencies are inferred from the claim text.