Lincoln Scott - Enzymatic synthesis and 19F NMR studies of 2-fluoroadenine-substituted RNA.

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  J Am Chem   Soc. 2004 Sep 29;126(38):11776-7.


  The production of isotopically labeled RNA remains critical to   current NMR structural studies. One approach to obtain simple NMR   spectra is to label with a nucleus that is not naturally   occurring in RNA. Fluorine-19 can serve as a sensitive   site-specific probe upon incorporation into RNA. Here we report   the efficient in vitro enzymatic synthesis of   2-fluoroadenosine-5'-triphosphate and its incorporation into the   HIV-2 transactivation region (TAR) of RNA by DNA   template-directed transcription using phage T7 RNA polymerase. We   provide unequivocal evidence for this 19F-substituted base   analogue capability to selectively interact with uracil, forming   2F-A-U base pairs in RNA. The introduction of a 2-fluoroadenyl   substitution is relatively nonperturbing and provides us with   uniquely positioned, sensitive NMR reporter groups to monitor   structural changes in the local RNA environment.

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