SEPARATION OF RNA PHOSPHOROTHIOATE OLIGONUCLEOTIDES BY HPLC

被引：42

作者：

Frederiksen, John K. ^{[1
,2
]}

Piccirilli, Joseph A. ^{[1
,3
]}

机构：

[1] Univ Chicago, Dept Biochem & Mol Biol, Chicago, IL 60637 USA

[2] Univ Chicago, Pritzker Sch Med, Chicago, IL 60637 USA

[3] Univ Chicago, Dept Chem, Chicago, IL 60637 USA

来源：

METHODS IN ENZYMOLOGY, VOL 468: BIOPHYSICAL, CHEMICAL, AND FUNCTIONAL PROBES OF RNA STRUCTURE, INTERACTIONS AND FOLDING, PT A | 2009年 / 468卷

关键词：

CATALYTIC METAL-ION; SOLID-PHASE SYNTHESIS; STEREOCHEMICAL COURSE; TETRAHYMENA RIBOZYME; HAMMERHEAD RIBOZYME; BINDING-SITE; FUNCTIONAL IDENTIFICATION; ACTIVE-SITE; P RNA; CLEAVAGE;

D O I：

10.1016/S0076-6879(09)68014-9

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Phosphorothioate oligonucleotides are indispensable tools for probing nucleic acid structure and function and for the design of antisense therapeutics. Many applications involving phosphorothioates require site- and stereospecific substitution of individual pro-R-p or pro-S-p nonbridging oxygens. However, the traditional approach to phosphorothioate synthesis produces a mixture of R-p and S-p diastereomers that must be separated prior to use. High-performance liquid chromatography (HPLC) has proven to be a versatile method for effecting this separation, with both reversed phase (RP) and strong anion exchange (SAX) protocols yielding favorable results. In this chapter, we present several examples of successful separations of RNA phosphorothioate diastereomers by HPLC. We also report the use of complementary DNA oligonucleotides for the separation of poorly resolved phosphorothioate RNAs.

引用

页码：289 / 309

页数：21

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