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  Site-specific labeling of RNA at internal ribose hydroxyl groups: Terbium-assisted deoxyribozymes at work.

Büttner, L., Javadi Zarnaghi, F., & Höbartner, C. (2014). Site-specific labeling of RNA at internal ribose hydroxyl groups: Terbium-assisted deoxyribozymes at work. Journal of the American Chemical Society, 135(22), 8131-8137. doi:10.1021/ja503864v.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0019-DBC9-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-3B09-5
Genre: Journal Article

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http://pubs.acs.org/doi/pdf/10.1021/ja503864v (Publisher version)
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Büttner, L.1, Author              
Javadi Zarnaghi, F.1, Author              
Höbartner, C.1, Author              
Affiliations:
1Research Group of Nucleic Acid Chemistry, MPI for Biophysical Chemistry, Max Planck Society, ou_578605              

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 Abstract: A general and efficient single-step method was established for site-specific post-transcriptional labeling of RNA. Using Tb3+ as accelerating cofactor for deoxyribozymes, various labeled guanosines were site-specifically attached to 2'-OH groups of internal adenosines in in vitro transcribed RNA. The DNA-catalyzed 2',5'-phosphodiester bond formation proceeded efficiently with fluorescent, spin-labeled, biotinylated, or cross-linker-modified guanosine triphosphates. The sequence context of the labeling site was systematically analyzed by mutating the nucleotides flanking the targeted adenosine. Labeling of adenosines in a purine-rich environment showed the fastest reactions and highest yields. Overall, practically useful yields >70% were obtained for 13 out of 16 possible nucleotide (nt) combinations. Using this approach, we demonstrate preparative labeling under mild conditions for up to similar to 160-nt-long RNAs, including spliceosomal U6 small nuclear RNA and a cyclic-di-AMP binding riboswitch RNA.

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Language(s): eng - English
 Dates: 2014-05-132014-06-04
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1021/ja503864v
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Title: Journal of the American Chemical Society
Source Genre: Journal
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Pages: - Volume / Issue: 135 (22) Sequence Number: - Start / End Page: 8131 - 8137 Identifier: -