| 摘要 |
A method is provided for determining the structure of a target RNA molecule in a living cell, comprising the steps of: (a) treating cells containing the target RNA with a base-modifying agent capable of disrupting Watson-Crick base pairing; (b) isolating RNA from treated cells; (c) hybridising the isolated RNA with a target RNA-specific first primer and reverse transcribing the target RNA molecules; and (d) assaying the length of the transcripts. The method allows the identification of nucleosides in the target RNA molecule which are exposed to attack by the base-modifying agents, since modification of a nucleoside will cause arrest of the reverse transcription reaction. Differences in exposure of particular nucleosides in an RNA molecule subjected to differing conditions in vivo represent differences in the secondary structure of the molecule, particularly differences in RNA:protein interactions. The method of the invention thus allows the comparison of the structure of RNA exposed to different influences. |
