New strategy for the synthesis of chemically modified RNA constructs exemplified by hairpin and hammerhead ribozymes

Proc Natl Acad Sci U S A. 2010 Aug 31;107(35):15329-34. doi: 10.1073/pnas.1006447107. Epub 2010 Aug 16.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chemistry / methods
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA / chemical synthesis*
  • RNA / chemistry
  • RNA, Catalytic / chemical synthesis*
  • RNA, Catalytic / chemistry

Substances

  • RNA, Catalytic
  • hairpin ribozyme
  • hammerhead ribozyme
  • RNA