Generalized RNA-directed recombination of RNA

Chem Biol. 2003 Dec;10(12):1233-43. doi: 10.1016/j.chembiol.2003.11.015.

Abstract

RNA strand exchange through phosphor-nucleotidyl transfer reactions is an intrinsic chemistry promoted by group I intron ribozymes. We show here that Tetrahymena and Azoarcus ribozymes can promote RNA oligonucleotide recombination in either two-pot or one-pot schemes. These ribozymes bind one oligonucleotide, cleave following a guide sequence, transfer the 3' portion of the oligo to their own 3' end, bind a second oligo, and catalyze another transfer reaction to generate recombinant oligos. Recombination is most effective with the Azoarcus ribozyme in a single reaction vessel in which over 75% of the second oligo can be rapidly converted to recombinant product. The Azoarcus ribozyme can also create a new functional RNA, a hammerhead ribozyme, which can be constructed via recombination and then immediately promote its own catalysis in a homogeneous milieu, mimicking events in a prebiotic soup.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Azoarcus / enzymology
  • Azoarcus / genetics
  • Base Pairing
  • Base Sequence
  • Catalysis
  • Introns / genetics
  • Kinetics
  • Molecular Sequence Data
  • RNA / chemistry
  • RNA / genetics*
  • RNA / metabolism*
  • RNA, Catalytic / chemistry
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • Recombination, Genetic / genetics*
  • Tetrahymena / enzymology
  • Tetrahymena / genetics

Substances

  • RNA, Catalytic
  • RNA