Molecular constructs for differentiating a target molecule from an off-target molecule
Molecular constructs, populations thereof, arrays, compositions, methods and kits for differentiating a target molecule from an off-target molecule are provided.
1. A population of two or more distinct molecular constructs capable of differentiating a target molecule from an off-target molecule and determining the target molecule concentration, said target molecule having a target domain and said off-target molecule having an off-target domain that differs from said target domain, each molecular construct comprising a distinct hybrid of
a) a first domain that is capable of binding to said target domain and to said off-target domain, and
b) a second domain that is at least partially hybridizable to said first domain,
wherein the first domain of each molecular construct of the population is identical;
wherein the free energy of displacement of the second domain from the first domain of the molecular construct by said target domain differs for each distinct molecular construct;
wherein for each distinct molecular construct the free energy of displacement of the second domain from the first domain by said target domain is more favorable than the free energy of displacement of the second domain from the first domain by the off-target domain;
wherein the second domain of each molecular construct of said population of molecular constructs comprises a first subdomain that is at least partially complementary to the first domain and a second subdomain that is not complementary to the first domain;
wherein the second subdomain of each molecular construct modulates the binding of the first domain to the second domain of each molecular construct; and
wherein
the second subdomain of the second domain of each molecular construct forms a G-tetraplex.
2. The population of claim 1 , wherein said off-target domain is generally homologous with respect to said target domain.
3. The population of claim 1 , wherein said off-target domain differs from said target domain either chemically by at least one functional group or conformationally or both.
4. The population of claim 1 , wherein said population comprises:
a) a first subpopulation comprising at least one said molecular construct with respect to a first off-target domain and
b) a second subpopulation comprising at least one said molecular construct with respect to a second off-target domain, and
wherein said first off-target domain differs from said second off-target domain.
5. The population of claim 4 , wherein said first off-target domain and said second off-target domain are disposed on the same off-target molecule.
6. The population of claim 4 , wherein said first off-target domain and said second off-target domain are disposed on different off-target molecules.
7. The population of claim 4 , wherein said first off-target domain is a target domain with respect to a second off-target molecule.
8. The population of claim 1 , wherein the first and second domains of each molecular construct are attached to a solid support.
9. The population of claim 1 , wherein each molecular construct is a nucleic acid molecule.
10. An array comprising more than one biosensor element on a solid support, wherein each biosensor element comprises a distinct molecular construct from the population of two or more distinct molecular constructs recited in claim 1 .
11. A kit comprising at least one population of molecular constructs of claim 1 .
12. An array of a plurality of populations in accordance with claim 1 for identifying a plurality of different target molecules in a sample and determining the relative number of each of those different target molecules in the sample,
wherein
the free energy of displacement from a molecular construct of one population for a target molecule is distinct from the free energy of displacement of other molecular constructs of the same or of another population for the same target molecule or for any other target molecule, such that each different target molecule in said sample can be identified, and the relative number of each of the different target molecules in said sample can be determined.
13. The array of claim 12 , wherein the number of different targets is from about 2 to 10.
14. The array of claim 12 , wherein the number of different targets is from 100 to 1000.
15. The array of claim 12 , wherein the number of different targets is greater than 1000.
16. An array of two or more populations in accordance with claim 1 , wherein the first domain of each molecular construct of a population is distinct from the first domain of each molecular construct of another population.
17. The array of claim 16 , wherein the target domain which binds the first domain of the molecular constructs of a population is disposed on the same target molecule as the target domain which binds the first domains of the molecular constructs of another population.
18. The array of claim 16 , wherein the target domain which binds the first domain of the molecular constructs of a population is an off-target domain for the first domain of the molecular construct of another population.
19. A method for differentiating a target molecule from an off-target molecule, said method comprising contacting the population of molecular constructs of claim 1 with an unknown molecule.