Synthesis of oligo-RNAs with photocaged adenosine 2'-hydroxyls.
This protocol describes a general method for the preparation of RNAs in which the reactivity or hydrogen-bonding properties of the molecule are modified in a photoreversible fashion by use of a caging strategy. A single caged adenosine, modified at the 2' position as a nitro-benzyl ether, can be incorporated into short RNAs by chemical synthesis or into long RNAs by a combination of chemical and enzymatic synthesis. The modified RNAs can be uncaged by photolysis under a variety of conditions including the use of a laser or xenon lamp, and the course of this uncaging reaction may be readily followed by HPLC or thin-layer chromatography.
Chaulk
Steven G
SG
Department of Biochemistry, University of Alberta Edmonton, Alberta, Canada T6G 2H7.
MacMillan
Andrew M
AM
D000241:Adenosine / Q000737:chemistry; D002627:Chemistry, Physical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006860:Hydrogen Bonding; D006878:Hydroxides / Q000737:chemistry; D010782:Photolysis; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry*
D006878:Hydroxides; D012313:RNA; C031356:hydroxide ion; D000241:Adenosine