Single-molecule selection methods and compositions therefrom
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12P-019/34
C07H-021/02
출원번호
US-0907385
(2001-07-17)
발명자
/ 주소
Cubicciotti, Roger S.
출원인 / 주소
Molecular Machines, Inc.
대리인 / 주소
Licata & Tyrrell P.C.
인용정보
피인용 횟수 :
34인용 특허 :
27
초록▼
Single-molecule selection methods are provided for identifying target-binding molecules from diverse sequence and shape libraries. Complexes and imprints of selected target-binding molecules are also provided. The subject selection methods are used to identify oligonucleotide and nonnucleotide molec
Single-molecule selection methods are provided for identifying target-binding molecules from diverse sequence and shape libraries. Complexes and imprints of selected target-binding molecules are also provided. The subject selection methods are used to identify oligonucleotide and nonnucleotide molecules with desirable properties for use in pharmaceuticals, drug discovery, drug delivery, diagnostics, medical devices, cosmetics, agriculture, environmental remediation, smart materials, packaging, microelectronics and nanofabrication. Single oligonucleotide molecules with desirable binding properties are selected from diverse sequence libraries and identified by amplification and sequencing. Alternatively, selected oligonucleotide molecules are identified by sequencing without amplification. Nonnucleotide molecules with desirable properties are identified by single-molecule selection from libraries of conjugated molecules or nucleotide-encoded nonnucleotide molecules. Alternatively, target-specific nonnucleotide molecules are prepared by imprinting selected oligonucleotide molecules into nonnucleotide molecular media. Complexes and imprints of molecules identified by single-molecule selection are shown to have broad utility as drugs, prodrugs, drug delivery systems, willfully reversible cosmetics, diagnostic reagents, sensors, transducers, actuators, adhesives, adherents and novel multimolecular devices.
대표청구항▼
1. A method of identifying an oligonucleotide molecule that specifically binds to a nonoligonucleotide molecule by sequence-dependent interaction, said method comprising:(a) contacting a mixture comprising oligonucleotide molecules with a nonoligonucleotide molecule that is not a biological recognit
1. A method of identifying an oligonucleotide molecule that specifically binds to a nonoligonucleotide molecule by sequence-dependent interaction, said method comprising:(a) contacting a mixture comprising oligonucleotide molecules with a nonoligonucleotide molecule that is not a biological recognition site for nucleic acids;(b) detecting with single-molecule resolution a single complex comprising an oligonucleotide molecule specifically bound by sequence-dependent interaction to said nonoligonucleotide molecule; and(c) determining, transcribing, amplifying or replicating a nucleotide sequence of said oligonucleotide molecule to identify the oligonucleotide molecule. 2. The method of claim 1 further comprising dissociating said complex prior to determining, transcribing, amplifying or replicating a nucleotide sequence of the oligonucleotide molecule. 3. The method of claim 1 wherein the oligonucleotide molecule comprises at least one of a conjugated molecule or a nucleotide-encoded nonnucleotide molecule. 4. A method of identifying an oligonucleotide molecule from a mixture of oligonucleotide molecules, said method comprising:(a) contacting a mixture of oligonucleotide molecules with a nonoligonucleotide molecule that is not a biological recognition site for nucleic acids;(b) detecting with single-molecule resolution a single complex comprising an oligonucleotide molecule specifically bound by sequence dependent interaction to said nonoligonucleotide molecule; and(c) identifying at least a portion of the oligonucleotide molecule by determining a nucleotide sequence, recognition property, binding specificity, binding force or chemical identity of the oligonucleotide molecule. 5. The method of claim 4 further comprising dissociating said complex prior to identifying at least a portion of the oligonucleotide molecule. 6. The method of claim 4 wherein the oligonucleotide molecule comprises at least one of a conjugated molecule or a nucleotide-encoded nonnucleotide molecule. 7. A method of identifying a nucleotide sequence comprising an oligonucleotide molecule that specifically binds by sequence dependent interaction to a selected target molecule immobilized on a solid support, said method comprising;(a) immobilizing a selected target molecule on a solid support;(b) contacting the immobilized target molecule with a mixture comprising oligonucleotide molecules;(c) detecting with single-molecule resolution a single complex comprising an oligonucleotide molecule specifically bound by sequence-dependent interaction to the immobilized target molecule; and(d) identifying a nucleotide sequence of the oligonucleotide molecule via sequence determination, transcription, amplification or replication. 8. The method of claim 7 further comprising dissociating said complex prior to identifying a nucleotide sequence of the oligonucleotide molecule. 9. The method of claim 7 wherein the oligonucleotide molecule comprises at least one of a conjugated molecule or a nucleotide-encoded nonnucleotide molecule. 10. A method of identifying without amplification a nucleotide sequence of an oligonucleotide molecule that specifically binds by sequence-dependent interaction to a selected target molecule, said method comprising;(a) contacting a mixture comprising an assortment of oligonucleotide molecules with a selected nonoligonucleotide target molecule that is not a biological recognition site for nucleic acids;(b) separating a target -bond oligonucleotide molecule from other oligonucleotide molecule of the mixture; and(c) identifying without amplification a nucleotide sequence of the target-bound oligonucleotide molecule. 11. The method of claim 10 further comprising dissociating said target-bound oligonucleotide molecule from its target prior to identifying a nucleotide sequence of the target-bound oligonucleotide molecule. 12. The method of claim 10 wherein said target-bound oligonucleotide molecule comprises at least one of a conjugated molecule or a n ucleotide encoded nonnucleotide molecule. 13. A method of detecting a single nonoligonucleotide molecule comprising:(a) selecting a nonnaturally occurring oligonucleotide molecule capable of specifically binding to a selected nonoligonucleotide molecule that is not a biological recognition site f or nucleic acids;(b) preparing a reaction mixture comprising said nonnaturally occurring oligonucleotide molecule and said selected nonoligonucleotide molecule; and(c) detecting with single-molecule resolution a single complex comprising said nonnaturally occurring oligonucleotide molecule specifically bound to said selected nonoligonucleotide molecule. 14. The method of claim 13 further comprising determining, transcribing, amplifying or replicating a nucleotide sequence of the specifically bound oligonucleotide molecule. 15. The method of claim 13 further comprising identifying at least a portion of the specifically bound oligonucleotide molecule by determining a nucleotide sequence, recognition property, binding specificity, binding force or chemical identity of the specifically bound oligonucleotide molecule. 16. The method of claim 13 wherein said nonnaturally occurring oligonucleotide molecule comprises at least one of a conjugated molecule or a nucleotide-encoded nonnucleotide molecule.
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