Detection of nucleic acid lesions and adducts using nanopores
Methods, systems, and compounds for detecting modified nucleic acid bases are disclosed and described. The methods provide for detecting a nucleic acid lesion and can include directing a nucleic acid adduct into a channel, wherein the nucleic acid adduct includes a nucleic acid having a lesion and a current modulating compound coupled to the nucleic acid at the lesion ( 110 ), and measuring a change in current through the channel in response to the current modulating compound to detect the lesion ( 112 ). The method can optionally include forming the nucleic acid adduct. Also provided is a method for identifying the number of repeat nucleotides in at least a portion of a nucleic acid strand, a method of assigning a registration marker within a nucleic acid, and a method of obtaining sequence information from a nucleic acid comprising assigning a registration marker on the nucleic acid.
1. A system for detecting a current modulating compound, comprising:
a membrane including an opening;
a pair of electrodes configured to register changes in electrical current across the opening; and
a nucleic acid comprising a nucleic acid adduct comprising a current modulating compound coupled to a nucleotide using a coupling reaction selected from the group consisting of oxidation, alkylation, platination, deamination, halogenation, glycosylation, conversion to an abasic site and further adduct formation, and combinations thereof, wherein the nucleic acid adduct is at least partially positioned within the opening, and wherein the current modulating compound is coupled to the nucleic acid at an abasic site.
2. The system of claim 1 , wherein the opening is a nanopore.
3. The system of claim 2 , wherein the nanopore is a conical nanopore.
4. The system of claim 1 , wherein a lipid bilayer is suspended across the opening and the lipid bilayer includes a protein embedded in the lipid bilayer to form a channel such that transport of the nucleic acid adduct across the channel is inhibited while transport of non-adduct nucleic acid is not substantially inhibited.
5. The system of claim 1 , wherein the current modulating compound includes a member selected from the group consisting of an alkane, an alkene, an alkyne, an aryl, a sugar, a carbohydrate, an azide, a halide, an amine, an imine, a peptide, a crown ether, a metal-binding ligand, a transition metal complex, and a combination thereof.
6. The nucleic acid of claim 1 , wherein the current modulating compound comprises a crown ether.
7. The nucleic acid of claim 6 , wherein the crown ether is selected from the group consisting of 18-crown-6 and 15-crown-5.
8. A system for detecting a current modulating compound, comprising:
a membrane including an opening;
a pair of electrodes configured to register changes in electrical current across the opening; and
a nucleic acid comprising a nucleic acid adduct comprising a current modulating compound coupled to a nucleotide using a coupling reaction selected from the group consisting of oxidation, alkylation, platination, deamination, halogenation, glycosylation, conversion to an abasic site and further adduct formation, and combinations thereof, wherein the nucleic acid adduct is at least partially positioned within the opening, wherein the current modulating compound comprises a crown ether.
9. The nucleic acid of claim 8 , wherein the crown ether is selected from the group consisting of 18-crown-6 and 15-crown-5.