Post-synthetically ligated ribozymes: an alternative approach to iterative solid-phase synthesis.
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.
Bellon
L
L
Department of Oligonucleotide Chemistry, Ribozyme Pharmaceuticals, Inc., Boulder, Colorado 80301, USA. lbellon@rpi.com
Workman
C T
CT
Jarvis
T C
TC
Wincott
F E
FE
D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D015195:Drug Design; D007202:Indicators and Reagents; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects; D009690:Nucleic Acid Conformation; D010084:Oxidation-Reduction; D016337:RNA, Catalytic / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D051381:Rats
D007202:Indicators and Reagents; D008024:Ligands; D016337:RNA, Catalytic