A general and efficient approach for the construction of RNA oligonucleotides containing a 5'-phosphorothiolate linkage.
A general and efficient approach for the construction of RNA oligonucleotides containing a 5'-phosphorothiolate linkage.
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DOI：
10.1093/nar/gkq1265
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发表时间：
2011-03
影响因子：
14.9
通讯作者：
Piccirilli JA
中科院分区：
文献类型：
--
作者：
Li NS;Frederiksen JK;Koo SC;Lu J;Wilson TJ;Lilley DM;Piccirilli JA
Oligoribonucleotides containing a 5′-phosphorothiolate linkage have provided effective tools to study the mechanisms of RNA catalysis, allowing resolution of kinetic ambiguity associated with mechanistic dissection and providing a strategy to establish linkage between catalysis and specific functional groups. However, challenges associated with their synthesis have limited wider application of these modified nucleic acids. Here, we describe a general semisynthetic strategy to obtain these oligoribonucleotides reliably and relatively efficiently. The approach begins with the chemical synthesis of an RNA dinucleotide containing the 5′-phosphorothiolate linkage, with the adjacent 2′-hydroxyl group protected as the photolabile 2′-O-o-nitrobenzyl or 2′-O-α-methyl-o-nitrobenzyl derivative. Enzymatic ligation of the 2′-protected dinucleotide to transcribed or chemically synthesized 5′ and 3′ flanking RNAs yields the full-length oligoribonucleotide. The photolabile protecting group increases the chemical stability of these highly activated oligoribonucleotides during synthesis and long-term storage but is easily removed with UV irradiation under neutral conditions, allowing immediate use of the modified RNA in biochemical experiments.
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影响因子：
4.5
作者：
Gordon, PM;Sontheimer, EJ;Piccirilli, JA
通讯作者：
Piccirilli, JA
影响因子：
14.9
作者：
BURGIN, AB;HUIZENGA, BN;NASH, HA
通讯作者：
NASH, HA
影响因子：
3.6
作者：
IYER, RP;PHILLIPS, LR;BEAUCAGE, SL
通讯作者：
BEAUCAGE, SL
影响因子：
14.9
作者：
Kuimelis, RG;McLaughlin, LW
通讯作者：
McLaughlin, LW
影响因子：
14.9
作者：
Chaulk, SG;MacMillan, AM
通讯作者：
MacMillan, AM