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Updated: Feb 11, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
Published on: May 3, 2014
Separation of Spliceosome Assembly from Catalysis with Caged pre-mRNA Substrates
Steven G Chaulk1, Andrew M MacMillan2
1Department of Chemistry University of Toronto Toronto, ON M5S 3H6 (Canada).
Abstract:
Getting spliced? Pre-mRNA splicing is catalyzed by the spliceosome, a complex assembly of proteins and RNA, which forms in an ordered fashion on the substrate. If one of the residues of the substrate is caged with a photolabile o-nitrobenzyl group, the splicing reaction can be transiently blocked, until subsequent initiation by photolysis of the complexes. This RNA-caging approach effectively separates the spliceosome assembly from the catalytic reaction and allows the two processes to be studied independently.
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