Abstract
Modified guanosine monophosphates have been employed to introduce various functional groups onto RNA 5'-ends. Applications of modified RNA 5'-ends include the generation of functionalized RNA libraries for in vitro selection of catalytic RNAs, the attachment of photoaffinity-tags for mapping RNA-protein interactions or active sites in catalytic RNAs, or the non-radioactive labeling of RNA molecules with fluorescent groups. While in these and in similar applications a stable linkage is desired, in selection experiments for generating novel catalytic RNAs it is often advantageous that a functional group is introduced reversibly. Here we give a quantitative comparison of the different strategies that can be applied to reversibly attach functional groups via disulfide bonds to RNA 5'-ends. We report the preparation of functional groups with disulfide linkages, their incorporation efficiency into an RNA library, and their stability under various conditions. Copyright (C) 2000 Elsevier Science Ltd.
Original language | English (US) |
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Pages (from-to) | 1317-1329 |
Number of pages | 13 |
Journal | Bioorganic and Medicinal Chemistry |
Volume | 8 |
Issue number | 6 |
DOIs | |
State | Published - Jun 2000 |
Bibliographical note
Funding Information:This study was supported by grants of the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie to M.F. and by a NATO travel grant to M. F. and J. S. N. We thank Nicole Rubner for operating HPLC-stability studies, Carol Zehetmeier for excellent technical assistance, Michael Blind, Günter Mayer, Felix Hausch, Barbara Gatto, Nikolai Raffler and Oliver Thum for helpful discussions and Wolfgang Steglich and Ernst-L. Winnacker for support. A. J. and B. S. express their gratitude to the DFG for support. J. S. N. thanks the following agencies for support in the form of awards: the Camille and Henry Dreyfus Foundation (Teacher-Scholar Award), the Alfred P. Sloan Foundation (Alfred P. Sloan Research Fellowship), and the American Chemical Society (Arthur C. Cope Scholar Award).