{"search_session":{},"preferences":{"l":"en","queryLanguage":"en"},"patentId":"US_9470680_B2","frontPageModel":{"patentViewModel":{"ref":{"entityRefId":"148-802-168-437-428","entityRefType":"PATENT"},"entityMetadata":{"linkedIds":{"empty":true},"tags":[],"collections":[{"id":8759,"type":"PATENT","title":"University of Columbia","description":"","access":"OPEN_ACCESS","displayAvatar":true,"attested":false,"itemCount":13487,"tags":[],"user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"notes":[{"id":8203,"type":"COLLECTION","user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"text":"
Searched applicants and Owners= \"Columbia Univ\", \"Univ Columbia\", \" Univ Colum*\", \"Colum* univ\", \"Univ* Colum* NOT British\".
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Searched applicants and Owners= \"Columbia Univ\", \"Univ Columbia\", \" Univ Colum*\", \"Colum* univ\", \"Univ* Colum* NOT British\".
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Total patents: more than 10k
b) mapping the phylogenetically-variable amino acids in the three-dimensional structure of the class 1 RF;\n
c) removing all wildtype cysteine residues;\n
d) replacing each of two phylogenetically-variable amino acids in an inactive region of the class 1 RF with a cysteine residue; and\n
e) conjugating two fluorescent labels to the cysteine residues; thereby making the class 1 RF conjugated to two fluorescent labels of claim 1."],"number":6,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 6, wherein the two fluorescent labels are a donor-acceptor pair."],"number":7,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 7, further comprising:\n
F) isolating class 1 RFs conjugated to a donor-acceptor pair."],"number":8,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 8, wherein the isolating is by hydrophobic interaction chromatography."],"number":9,"annotation":false,"claim":true,"title":false},{"lines":["A method of detecting a conformational change in a class 1 RF, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface;\n
b) adding to the reaction mixture a probe comprising the class 1 RF conjugated to a donor-acceptor pair of fluorescent labels of claim 3; and\n
c) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair;\nwherein a shift in FRET efficiency after adding the probe indicates a conformational change in the class 1 RF."],"number":10,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 10, wherein the donor-acceptor pair is Cy3/Cy5."],"number":11,"annotation":false,"claim":true,"title":false},{"lines":["A method of assaying RF3 activity, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface;\n
b) adding to the reaction mixture a probe comprising a class 1 RF conjugated to the donor-acceptor pair of fluorescent labels of claim 3;\n
c) adding to the reaction mixture RF3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imaging the solid surface;\nwherein a loss in FRET efficiency after adding the RF3 indicates RF3 activity by indicating release of the class 1 RF from the ribosome complex."],"number":12,"annotation":false,"claim":true,"title":false},{"lines":["A method of assaying RF3 activity, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface;\n
b) adding to the reaction mixture a probe comprising a class 1 RF conjugated to the donor-acceptor pair of fluorescent labels of claim 3;\n
c) adding to the reaction mixture RF3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imagining the reaction mixture;\nwherein high FRET efficiency after adding the RF3 indicates RF3 activity by indicating release of the class 1 RF from the ribosome complex."],"number":13,"annotation":false,"claim":true,"title":false},{"lines":["A method of identifying a compound for reducing nonsense-mediated decay of mRNA, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface, wherein the reaction mixture comprises an mRNA comprising a premature stop codon;\n
b) adding to the reaction mixture a candidate compound;\n
c) adding to the reaction mixture a probe comprising a class 1 RF conjugated to the donor-acceptor pair of fluorescent labels of claim 3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imaging the reaction mixture;\nwherein high FRET efficiency after adding the probe indicates that the candidate compound inhibits binding of the class 1 RF to the ribosome complex, thereby identifying a compound for reducing nonsense-mediated decay of mRNA."],"number":14,"annotation":false,"claim":true,"title":false},{"lines":["A method of identifying a compound for reducing nonsense-mediated decay of mRNA, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface, wherein the reaction mixture comprises an mRNA comprising a premature stop codon;\n
b) adding to the reaction mixture a candidate compound;\n
c) adding to the reaction mixture a probe comprising the class 1 RF conjugated to a donor-acceptor pair of fluorescent labels of claim 3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imaging the solid surface;\nwherein lack of FRET efficiency or transient FRET efficiency after adding the probe indicates that the candidate compound inhibits binding of the class 1 RF to the ribosome complex, thereby identifying a compound for reducing nonsense-mediated decay of mRNA."],"number":15,"annotation":false,"claim":true,"title":false},{"lines":["A method of identifying a compound for inhibiting termination of protein synthesis at a premature stop codon, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface, wherein the ribosome complex comprises a first fluorescent label, and wherein the reaction mixture comprises an mRNA comprising a premature stop codon;\n
b) adding to the reaction mixture a candidate compound;\n
c) adding to the reaction mixture a probe comprising the class 1 RF conjugated to a donor-acceptor pair of fluorescent labels of claim 3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imaging the reaction mixture;\nwherein high FRET efficiency after adding the probe indicates that the candidate compound inhibits binding of the class 1 RF to the ribosome complex, thereby identifying a compound for inhibiting termination of protein synthesis at a premature stop codon."],"number":16,"annotation":false,"claim":true,"title":false},{"lines":["A method of identifying a compound for inhibiting termination of protein synthesis at a premature stop codon, the method comprising:\n
a) providing a reaction mixture comprising an isolated, translationally-competent bacterial ribosome complex bound to a solid surface, wherein the ribosome complex comprises a first fluorescent label, and wherein the reaction mixture comprises an mRNA comprising a premature stop codon;\n
b) adding to the reaction mixture a candidate compound;\n
c) adding to the reaction mixture a probe comprising a class 1 RF conjugated to the donor-acceptor pair of fluorescent labels of claim 3; and\n
d) measuring fluorescence resonance energy transfer (FRET) efficiency between the donor-acceptor pair by imaging the solid surface;\nwherein lack of FRET efficiency or transient FRET efficiency after adding the probe indicates that the candidate compound inhibits binding of the class 1 RF to the ribosome complex, thereby identifying a compound for inhibiting termination of protein synthesis at a premature stop codon."],"number":17,"annotation":false,"claim":true,"title":false}]}},"filters":{"npl":[],"notNpl":[],"applicant":[],"notApplicant":[],"inventor":[],"notInventor":[],"owner":[],"notOwner":[],"tags":[],"dates":[],"types":[],"notTypes":[],"j":[],"notJ":[],"fj":[],"notFj":[],"classIpcr":[],"notClassIpcr":[],"classNat":[],"notClassNat":[],"classCpc":[],"notClassCpc":[],"so":[],"notSo":[],"sat":[]},"sequenceFilters":{"s":"SEQIDNO","d":"ASCENDING","p":0,"n":10,"sp":[],"si":[],"len":[],"t":[],"loc":[]}}