Continuous in vitro evolution of catalytic function.

A population of RNA molecules that catalyze the template-directed ligation of RNA substrates was made to evolve in a continuous manner in the test tube. A simple serial transfer procedure was used to achieve approximately 300 successive rounds of catalysis and selective amplification in 52 hours. During this time, the population size was maintained against an overall dilution of 3 x 10(298). Both the catalytic rate and amplification rate of the RNAs improved substantially as a consequence of mutations that accumulated during the evolution process. Continuous in vitro evolution makes it possible to maintain laboratory "cultures" of catalytic molecules that can be perpetuated indefinitely.

[1]  D. Mills,et al.  An extracellular Darwinian experiment with a self-duplicating nucleic acid molecule. , 1967, Proceedings of the National Academy of Sciences of the United States of America.

[2]  O. Uhlenbeck,et al.  Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. , 1987, Nucleic acids research.

[3]  M. Mörl,et al.  New reactions catalyzed by a group II intron ribozyme with RNA and DNA substrates , 1992, Cell.

[4]  G. F. Joyce,et al.  Directed evolution of an RNA enzyme. , 1992, Science.

[5]  AC Tose Cell , 1993, Cell.

[6]  D. Bartel,et al.  Isolation of new ribozymes from a large pool of random sequences [see comment]. , 1993, Science.

[7]  G. F. Joyce,et al.  Evolutionary optimization of the catalytic properties of a DNA-cleaving ribozyme. , 1994, Biochemistry.

[8]  R R Breaker,et al.  Emergence of a replicating species from an in vitro RNA evolution reaction. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A D Ellington,et al.  Artificial evolution and natural ribozymes , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  R R Breaker,et al.  Are engineered proteins getting competition from RNA? , 1996, Current opinion in biotechnology.

[11]  David P. Bartel,et al.  RNA-catalysed RNA polymerization using nucleoside triphosphates , 1996, Nature.

[12]  J W Szostak,et al.  Ribozymes: aiming at RNA replication and protein synthesis. , 1996, Chemistry & biology.

[13]  References and Notes , .