Diagnosing genetic disorders
First Claim
1. A method, comprising:
- a) providing a tRNA molecule, nucleic acid derived from a biological sample, and a marker;
b) aminoacylating said tRNA molecule with said marker to create a misaminoacylated tRNA;
c) introducing said misaminoacylated tRNA and said nucleic acid derived from a biological sample into a translation system under conditions such that said marker is incorporated into a nascent protein; and
d) screening for disease, said screening comprising detecting said marker.
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Abstract
This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as nucleic acids from biological samples, cells from bodily fluids, and nascent proteins from translation reactions. Agents comprise a detectable moiety bound to a photoreactive moiety. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed. The invention also relates to targets isolated with these conjugates which may be useful as pharmaceutical agents or compositions that can be administered to humans and other mammals. Useful compositions include biological agents such as nucleic acids, proteins, lipids and cytokines. Conjugates can also be used to monitor the pathway and half-life of pharmaceutical composition in vivo and for diagnostic, therapeutic and prophylactic purposes. The invention also relates to kits comprised of agents and conjugates that can be used for the detection of diseases, disorders and nearly any individual substance in a complex background of substances.
16 Citations
36 Claims
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1. A method, comprising:
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a) providing a tRNA molecule, nucleic acid derived from a biological sample, and a marker;
b) aminoacylating said tRNA molecule with said marker to create a misaminoacylated tRNA;
c) introducing said misaminoacylated tRNA and said nucleic acid derived from a biological sample into a translation system under conditions such that said marker is incorporated into a nascent protein; and
d) screening for disease, said screening comprising detecting said marker. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method of screening for a genetic disorder, comprising:
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a) providing a tRNA molecule, nucleic acid derived from a biological sample, and a detectable label;
b) aminoacylating said tRNA molecule with said detectable label to create a misaminoacylated tRNA;
c) introducing said misaminoacylated tRNA and said nucleic acid derived from a biological sample into a translation system under conditions such that said detectable label is incorporated into a nascent protein; and
d) screening for a genetic disorder, said screening comprising detecting said detectable label. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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Specification