Polymerase-Mediated Site-Specific Incorporation of a Synthetic Fluorescent Isomorphic G Surrogate into RNA.

ABSTRACT: An enzyme-mediated approach for the assembly of singly modified RNA constructs in which specific G residues are replaced with ^th G, an emissive isomorphic G surrogate, is reported. Transcription in the presence of ^th G and native nucleoside triphosphates enforces initiation with the unnatural analogue, yielding 5'-end modified transcripts that can be mono-phosphorylated and ligated to provide longer site-specifically modified RNA constructs. The scope of this unprecedented enzymatic approach to non-canonical purine-containing RNAs is explored via the assembly of several altered hammerhead (HH) ribozymes and a singly modified HH substrate. By strategically modifying key positions, a mechanistic insight into the ribozyme-mediated cleavage is gained. Additionally, the em

SUBMITTER: Li Y 

PROVIDER: S-EPMC5241218 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature