New strategy for the synthesis of chemically modified RNA constructs exemplified by hairpin and hammerhead ribozymes.
The CuAAC reaction (click chemistry) has been used in conjunction with solid-phase synthesis to produce catalytically active hairpin ribozymes around 100 nucleotides in length. Cross-strand ligation through neighboring nucleobases was successful in covalently linking presynthesized RNA strands with high efficiency (trans-ligation). In an alternative strategy, intrastrand click ligation was employed to produce a functional hammerhead ribozyme containing a novel nucleic acid backbone mimic at the catalytic site (cis-ligation). The ability to synthesize long RNA strands by a combination of solid-phase synthesis and click ligation is an important addition to RNA chemistry. It is compatible with a plethora of site-specific modifications and is applicable to the synthesis of many biologically important RNA molecules.
El-Sagheer
Afaf H
AH
School of Chemistry, University of Southampton, Highfield, Southampton, SO17 1BJ, United Kingdom.
Brown
Tom
T
D001483:Base Sequence; D002621:Chemistry / Q000379:methods; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry; D016337:RNA, Catalytic / Q000138:chemical synthesis* / Q000737:chemistry
D016337:RNA, Catalytic; C418481:hairpin ribozyme; C427204:hammerhead ribozyme; D012313:RNA