METHODS AND COMPOSITIONS FOR PROCESSES OF RAPID SELECTION AND PRODUCTION OF NUCLEIC ACID APTAMERS
First Claim
1. A composition of nucleic acid sequences comprising:
- a reporter moiety bound to a first strand of the nucleic acid sequence wherein the reporter moiety is capable of generating a signal; and
a signal reducing moiety bound to a second complimentary strand of the first nucleic acid sequence strand wherein the signal reducing moiety is capable of reducing the signal produced by the reporter moiety.
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Accused Products
Abstract
Embodiments herein relate to compositions and methods for making and using aptamers, for example, DNA aptamers (DCEs) and/or RNA aptamers. In some embodiments, methods relate to making and amplifying target DCEs. In certain embodiments, methods for making capture elements or aptamers concern using a reporter moiety and signal reducing moiety prior to amplifying a target-specific capture element. In some embodiments, methods disclosed herein may be used to rapidly generate large quantities of aptamers such as DCEs directed to a particular target agent. Some embodiments relate to systems for performing automated generation of aptamers.
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Citations
37 Claims
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1. A composition of nucleic acid sequences comprising:
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a reporter moiety bound to a first strand of the nucleic acid sequence wherein the reporter moiety is capable of generating a signal; and a signal reducing moiety bound to a second complimentary strand of the first nucleic acid sequence strand wherein the signal reducing moiety is capable of reducing the signal produced by the reporter moiety. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for producing nucleic acid sequences for isolating DNA aptamers (DCEs) directed to bind a target agent comprising:
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a) contacting a first reagent moiety with a first nucleic acid sequence mixture to form a first reagent moiety-nucleic acid sequence complex comprising nucleic acid sequences attached to first reagent moieties; and b) contacting a second reagent moiety with a second nucleic acid sequence mixture to form a second reagent moiety-nucleic acid sequence complex comprising nucleic acid sequences attached to second reagent moieties; wherein the first reagent moiety comprises a reporter moiety that is capable of generating a signal or a signal reducing moiety that is capable of reducing the signal from the reporter moiety and the second reagent moiety comprises the other of a reporter moiety or a signal reducing moiety. - View Dependent Claims (14, 15, 16)
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17. A method for producing nucleic acid aptamers comprising:
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a) contacting a first reagent moiety with a first nucleic acid sequence mixture to form a first moiety-nucleic acid sequence complex comprising a nucleic acid that is attached to the first reagent moiety; and b) contacting a second reagent moiety with a second nucleic acid sequence mixture to form a second moiety-nucleic acid sequence complex comprising a nucleic acid that is attached to the second reagent moiety wherein at least some of the nucleic acid sequences are complimentary to first nucleic acid sequences, wherein the first reagent moiety comprises a qdot moiety that is capable of generating a signal or a quenching agent moiety that is capable of absorbing the signal from the qdot and the second reagent moiety comprises the other of a qdot moiety or a quenching agent moiety. c) mixing the first moiety-nucleic acid sequence complex with the second moiety-nucleic acid sequence complex and allowing the formation of dsDNA (double-stranded DNA) complexes wherein the signal of the qdot moiety on one strand of the dsDNA is reduced when a quenching moiety is attached to a complimentary second strand on the dsDNA; d) introducing a target agent to the dsDNA complexes, wherein the target agent binds to at least the first strand or the second strand of the dsDNA complex to form a target agent-single-stranded DCE (DNA capture element) complex (target agent-ssDCE complex); and e) separating at least a portion of the target agent-ssDCE complex from the remainder of the complexes. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24)
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25. A system for producing DNA aptamers comprising:
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a) an element for inputting a first moiety reagent with a first mixture of nucleic acid sequences to form a first moiety reagent-nucleic acid sequence complex in a reaction vessel; and b) an element for inputting a second moiety reagent with a second mixture of nucleic acid sequences to form a second moiety reagent-nucleic acid sequence complex in another reaction vessel; wherein the first moiety reagent comprises a reporter moiety or a signal reducing moiety and the second reagent comprises the other of a reporter moiety or a signal reducing moiety. - View Dependent Claims (26, 27, 28, 29, 30)
f) an element for amplifying the single-stranded nucleic acid of the target agent-single-stranded nucleic acid complex bound to the solid substrate.
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31. A composition comprising, a quenching agent moiety bound to a dsDNA sequence wherein adsorption wavelength of the quenching agent moiety overlaps with emission wavelength of an agent capable of generating a signal.
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32. A kit for making aptamers comprising:
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a reporter moiety composition; and a signal reducing moiety composition. - View Dependent Claims (33, 34, 35, 36, 37)
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Specification