Journal Article10.1126/SCIENCE.1496376
Directed evolution of an RNA enzyme
Amber A. Beaudry,Gerald F. Joyce +1 more
TL;DR: An in vitro evolution procedure was used to obtain RNA enzymes with a particular catalytic function, and selection constraint was imposed on the population of ribozyme variants such that only those individuals that carried out DNA cleavage under physiologic conditions were amplified to produce "progeny" ribozymes.
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Abstract: An in vitro evolution procedure was used to obtain RNA enzymes with a particular catalytic function. A population of 10(13) variants of the Tetrahymena ribozyme, a group I ribozyme that catalyzes sequence-specific cleavage of RNA via a phosphoester transfer mechanism, was generated. This enzyme has a limited ability to cleave DNA under conditions of high temperature or high MgCl2 concentration, or both. A selection constraint was imposed on the population of ribozyme variants such that only those individuals that carried out DNA cleavage under physiologic conditions were amplified to produce "progeny" ribozymes. Mutations were introduced during amplification to maintain heterogeneity in the population. This process was repeated for ten successive generations, resulting in enhanced (100 times) DNA cleavage activity.
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Citations
Patent
Method for target site selection and discovery
James D. Thompson
- 08 Jul 1998
TL;DR: In this article, a method of screening/selection for nucleic acid catalysts, synthesis of ribozyme libraries and discovery of gene sequences involved in a biological process are described.
18
Replicability and Recurrence in the Experimental Evolution of a Group I Ribozyme
TL;DR: The diversity of solutions that result when the process of experimental evolution is repeated multiple times in a simple, engineered system is uncovered and underscores the complex network of possible trajectories through sequence space available to an evolving population.
Catalytic DNAzymes: derivations and functions.
TL;DR: It seems unlikely that CDzs will have clinical utility, but they are likely to have significant potential in the sensor/nanotechnology arena.
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Early Replicons: Origin and Evolution
Peter Schuster,Peter F. Stadler +1 more
- 01 Jan 2008
TL;DR: A theoretical account of molecular replication making precise the conditions under which one observes parabolic, exponential, or hyperbolic growth is presented in this paper, where computer simulations are used to model the interplay between adaptive evolution and random drift. Error propagation to forthcoming generations is analyzed in the absence of selective by neutral mutants as well as for predefined degrees of neutrality.
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Selection of Functional RNA and DNA Molecules from Randomized Sequences
Michael Famulok,Jack W. Szostak +1 more
- 01 Jan 1993
TL;DR: In vitro selection is a technique that allows the rapid screening of very large numbers of different DNA or RNA sequences for specific functional properties and the de novo isolation and evaluation of nucleic acids that bind to small biological and abiotic organic molecules.
17
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