Generalized RNA-directed recombination of RNA.
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.
Riley
Craig A
CA
Department of Chemistry, Portland State University, PO Box 751, Portland, OR 97207, USA.
Lehman
Niles
N
D000818:Animals; D020627:Azoarcus / Q000201:enzymology / Q000235:genetics; D020029:Base Pairing; D001483:Base Sequence; D002384:Catalysis; D007438:Introns / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics*; D013768:Tetrahymena / Q000201:enzymology / Q000235:genetics
D016337:RNA, Catalytic; D012313:RNA