Abstract
The trans insertion-splicing reaction, catalyzed by a group I intron-derived from Pneumocystis carinii, was recently developed for the site-specific insertion of a segment of RNA into a separate RNA substrate. The molecular determinants of this reaction for binding and catalysis are reasonably well understood, making them easily and highly modifiable for altering substrate specificity. To demonstrate proof-of-concept, we now report that the P. carinii ribozyme can except modified oligonucleotides as substrates for catalyzing the trans insertion-splicing reaction. Oligonucleotides that contain one or more sugar modifications (deoxy or methoxy substitution), a backbone modification (phosphorothioate substitution), or a base modification (2-aminopurine or 4-thiouridine) are effective substrates in this reaction. Apparently, trans insertion-splicing is a unique and viable reaction for the site-specific incorporation of modified oligonucleotides into RNAs. This is the first report of a group I intron-derived ribozyme being capable of catalyzing the insertion of a modified oligonucleotide into RNA.
Original language | English |
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Pages (from-to) | 81-84 |
Number of pages | 4 |
Journal | Archives of Biochemistry and Biophysics |
Volume | 478 |
Issue number | 1 |
DOIs | |
State | Published - Oct 1 2008 |
Bibliographical note
Funding Information:This research was supported by grants from the Kentucky Lung Cancer Research Program and The Lexington Foundation.
Keywords
- RNA modifications
- Ribozyme
- Trans insertion-splicing
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology