Nucleic acid mediated electron transfer
First Claim
1. A single-stranded nucleic acid containing at least one electron donor moiety and at least one electron acceptor moiety, said electron donor moiety and said electron acceptor moiety being covalently attached to said nucleic acid.
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Abstract
The present invention provides for the selective covalent modification of nucleic acids with redox active moieties such as transition metal complexes. Electron donor and electron acceptor moieties are covalently bound to the ribose-phosphate backbone of a nucleic acid at predetermined positions. The resulting complexes represent a series of new derivatives that are bimolecular templates capable of transferring electrons over very large distances at extremely fast rates. These complexes possess unique structural features which enable the use of an entirely new class of bioconductors and photoactive probes.
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Citations
20 Claims
- 1. A single-stranded nucleic acid containing at least one electron donor moiety and at least one electron acceptor moiety, said electron donor moiety and said electron acceptor moiety being covalently attached to said nucleic acid.
- 5. A composition comprising a first single stranded nucleic acid containing at least one electron donor moiety and a second single stranded nucleic acid containing at least one electron acceptor moiety, wherein said electron donor moiety and electron acceptor moiety are covalently linked to the ribose-phosphate backbone of said first and second single stranded nucleic acids.
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16. A method for making a single stranded nucleic acid containing an electron transfer moiety at the 5′
- terminus, comprising
a) incorporating a modified nucleotide into a growing nucleic acid at the 5′
position to form a modified single stranded nucleic acid;
b) hybridizing said modified single stranded nucleic acid with a complementary single stranded nucleic acid to form a double stranded nucleic acid;
c) reacting said double stranded nucleic acid with an electron transfer moiety such that said moiety is covalently attached to said modified single stranded nucleic acid; and
d) separating said complementary single stranded nucleic acid from said modified single stranded nucleic acid containing said electron transfer moiety.
- terminus, comprising
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17. A method for making a single stranded nucleic acid containing an electron transfer moiety covalently attached to an internal nucleotide, comprising
a) incorporating a modified nucleotide dimer into a growing nucleic acid to form a modified single stranded nucleic acid; -
b) hybridizing said modified single stranded nucleic acid with a complementary single stranded nucleic acid to form a double stranded nucleic acid;
c) reacting said double stranded nucleic acid with an electron transfer moiety such that said moiety is covalently attached to said modified single stranded nucleic acid; and
d) separating said complementary single stranded nucleic acid from the modified single stranded nucleic acid containing said electron transfer moiety.
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18. A method for making a single stranded nucleic acid containing an electron transfer moiety covalently attached to the 3′
- terminal nucleotide, comprising
a) incorporating a modified nucleotide via enzymatic addition or replacement into a nucleic acid;
b) hybridizing said modified single stranded nucleic acid with a complementary single stranded nucleic acid to form a double stranded nucleic acid;
c) reacting said double stranded nucleic acid with an electron transfer moiety such that said moiety is covalently attached through said phosphoramide bond of said modified single stranded nucleic acid; and
d) separating said complementary single stranded nucleic acid from the modified single stranded nucleic acid containing said electron transfer moiety.
- terminal nucleotide, comprising
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19. A method of detecting a target sequence in a nucleic acid sample comprising
a) hybridizing a single stranded nucleic acid containing at least one covalently attached electron donor moiety and at least one covalently attached electron acceptor moiety to said target sequence to form a hybridization complex; -
b) determining the electron transfer rate between said electron donor moiety and said electron acceptor moiety in the hybridization complex; and
c) comparing said electron transfer rate with the electron transfer rate in the absence of the target sequence as an indicator of the presence or absence of said target sequence.
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20. A method of detecting a target sequence in a nucleic acid wherein said target sequence comprises a first target domain and a second target domain adjacent to said first target domain, wherein said method comprises:
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a) hybridizing a first nucleic acid containing at least one electron donor moiety to said first target domain;
b) hybridizing a second nucleic acid containing at least one electron acceptor moiety to said second target domain;
c) determining the electron transfer rate between said electron donor moiety and said electron acceptor moiety while said first and second nucleic acids are hybridized to said first and second target domains; and
d) comparing said electron transfer rate with the electron transfer rate in the absence of the target sequence as an indicator of the presence or absence of said target sequence in said nucleic acid sample.
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Specification