Methods for identifying RNA segments bound by RNA-binding proteins or ribonucleoprotein complexes

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United States of America Patent

PATENT NO 8841073
APP PUB NO 20110287412A1
SERIAL NO

13055976

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Abstract

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The present invention relates to a method for identifying a binding site on an RNA transcript, wherein the binding site binds to one or more binding moieties. The method includes, among other things, introducing a photoreactive nucleoside into living cells wherein the living cells incorporate the photoreactive nucleoside into RNA transcripts during transcription thereby producing modified RNA transcripts; reverse transcribing the RNA of isolated cross-linked segments thereby generating cDNA transcripts with one mutation wherein the photoreactive nucleoside is transcribed to a mismatched deoxynucleoside; amplifying the cDNA transcripts thereby generating amplicons; and analyzing the sequences of the amplicons aligned against the reference sequence so as to identify the binding site, wherein the sequences of each amplicon having a mutation resulting from the introduction of the photoreactive nucleoside is considered to be a valid amplicon comprising at least a portion of a binding site on the RNA transcript.

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Patent Owner(s)

Patent OwnerAddress
UNIVERSITY OF BASELVIZEREKTORAT FORSCHUNG PETERSGRABEN 35 BASEL CH-4001

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Ascano,, Jr Manuel New YorK, US 7 127
Burger, Lukas Basel, CH 2 8
Hafner, Markus New York, US 7 22
Khorshid, Mohsen Basel, CH 2 8
Landthaler, Markus Berlin, DE 11 275
Tuschl, Thomas Brooklyn, US 86 5967
Zavolan, Mihaela Binningen, CH 2 8

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