Post-synthetically Ligated Ribozymes:  An Alternative Approach to Iterative Solid-Phase Synthesis

• Published in: Bioconjugate chemistry Vol. 8; no. 2; pp. 204 - 212
• Main Authors: Bellon, Laurent, Workman, Christopher T, Jarvis, Thale C, Wincott, Francine E
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.03.1997
• Subjects: Animals; Base Sequence; Cell Division - drug effects; Cells, Cultured; Drug Design; Indicators and Reagents; Kinetics; Ligands; Molecular Sequence Data; Molecular Structure; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Nucleic Acid Conformation; Oxidation-Reduction; Rats; RNA, Catalytic - chemical synthesis; RNA, Catalytic - chemistry; RNA, Catalytic - metabolism
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc970011q

To improve the overall yield of ribozyme synthesis, a convergent approach, based on the post-synthetic formation of an amino linker between two half-ribozymes was investigated. Borane·pyridine-mediated reductive amination of 3‘-phosphoglycaldehyde-5‘-half-ribozymes with 5‘-aminohexyl-3‘-half-ribozymes generated the corresponding amino-linked ribozymes in yields >77% on different scales. The investigation of a variety of reducing agents is discussed together with a kinetic analysis of the selected coupling reaction. These post-synthetically ligated ribozymes exhibited slightly reduced in vitro catalytic activity and cell efficacy.