Nucleic acid based fluorescent sensor for copper detection
View Patent ↗A nucleic acid enzyme responsive to copper, comprising an oligonucleotide comprising a nucleotide sequence of SEQ ID NO:1, wherein the nucleic acid enzyme is not self-cleaving.
1. An isolated nucleic acid enzyme activated in the presence of copper, comprising an oligonucleotide comprising a nucleotide sequence shown in SEQ ID NO: 1, wherein the nucleic acid enzyme is not self-cleaving.
2. The nucleic acid enzyme of claim 1 , wherein the oligonucleotide further comprises a first quencher at a 5′-end or a 3′-end of the nucleic acid enzyme, and optionally further comprises a spacer between the oligonucleotide and the first quencher.
3. The nucleic acid enzyme of claim 2 , wherein the oligonucleotide comprises a nucleotide sequence shown in SEQ ID NO:5.
4. An isolated nucleic acid enzyme activated in the presence of copper, comprising:
an oligonucleotide comprising a nucleotide sequence of SEQ ID NO:1, and
a quencher, attached at a 3′ end of the nucleic acid enzyme.
5. The nucleic acid enzyme of claim 1 , wherein the oligonucleotide further comprises a first quencher at a 3′-end of the nucleic acid enzyme, and a spacer between the oligonucleotide and the first quencher.
6. The nucleic acid enzyme of claim 2 , wherein the oligonucleotide comprises a first quencher at a 5′-end of the nucleic acid enzyme.
7. A sensor system for determining an amount of copper in a sample, comprising:
the nucleic acid enzyme of claim 2 ,
a substrate for the nucleic acid enzyme,
a second quencher, attached to the substrate, and
a fluorophore, attached to the substrate.
8. A sensor system for determining an amount of copper in a sample, comprising:
the nucleic acid enzyme of claim 5 ,
a substrate for the nucleic acid enzyme,
a second quencher, attached to the substrate, and
a fluorophore, attached to the substrate.
9. A sensor system for determining an amount of copper in a sample, comprising:
the nucleic acid enzyme of claim 6 ,
a substrate for the nucleic acid enzyme,
a second quencher, attached to the substrate, and
a fluorophore, attached to the substrate.
10. The sensor system of claim 7 , wherein the substrate comprises an oligonucleotide comprising a nucleotide sequence shown in SEQ ID NO:4, wherein the second quencher is at a 5′-end of the substrate, the fluorophore is at a 3′-end of the substrate, wherein the substrate optionally further comprises a spacer between the oligonucleotide and the second quencher, and wherein the substrate optionally further comprises a spacer between the oligonucleotide and the fluorophore.
11. The sensor system of claim 8 , wherein the substrate comprises an oligonucleotide comprising a nucleotide sequence shown in SEQ ID NO:4, wherein the second quencher is at a 5′-end of the substrate, the fluorophore is at a 3′-end of the substrate, wherein the substrate optionally further comprises a spacer between the oligonucleotide and the second quencher, and wherein the substrate optionally further comprises a spacer between the oligonucleotide and the fluorophore.
12. The sensor system of claim 7 , wherein the substrate comprises an oligonucleotide comprising a nucleotide sequence shown in SEQ ID NO:6, wherein the second quencher is at a 5′-end of the substrate, the fluorophore is at a 3′-end of the substrate, wherein the substrate optionally further comprises a spacer between the oligonucleotide and the second quencher, and wherein the substrate optionally further comprises a spacer between the oligonucleotide and the fluorophore.
13. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 7 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
14. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 8 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
15. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 9 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
16. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 10 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
17. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 11 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
18. A method of measuring an amount of copper in a sample, comprising:
mixing the sample with the sensor system of claim 12 ; and
detecting fluorescence, wherein detecting an increase in fluorescence indicates the presence of copper.
19. The sensor system of claim 7 , wherein the fluorophore is 6-carboxyfluorescein and the first and second quencher are Iowa Black FQ.
20. The method of claim 13 , wherein the sample is further mixed with ascorbate.