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Nucleic Acids Research 2006 34(3):e21; doi:10.1093/nar/gnj018
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Published online 10 February 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

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Isolation of a site-specifically modified RNA from an unmodified transcript

Ya-Ming Hou*, Zhi Li and Howard Gamper

Department of Biochemistry and Molecular Biology, Thomas Jefferson University 233 South 10th Street, BLSB 220, Philadelphia, PA 19107-5719, USA

*To whom correspondence should be addressed. Tel: +1 215 503 4480; Fax: +1 215 503 4954; Email: Ya-Ming.Hou{at}jefferson.edu

Received November 20, 2005. Revised January 8, 2006. Accepted January 19, 2006.

Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies.


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