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
Catalytic rna molecules
Gerald F. Joyce
- 16 Jul 1997
TL;DR: In this paper, the authors present a set of enzymes capable of cleaving nucleic acid molecules, including single-stranded DNA, in a site-specific manner under physiologic conditions, as well as compositions including same.
19
Design and Experimental Evolution of trans-Splicing Group I Intron Ribozymes
TL;DR: This review provides an overview of the different types of trans-splicing group I intron ribozymes that have been generated, and the experimental evolution systems that have be used to improve them.
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A Synthetic Protein Selected for Ligand Binding Affinity Mediates ATP Hydrolysis
Chad R. Simmons,Joshua M. Stomel,Michael D. McConnell,Daniel A. Smith,Jennifer L. Watkins,James P. Allen,John C. Chaput +6 more
TL;DR: It is shown that a de novo evolved ATP binding protein, selected solely on the basis of its ability to bind ATP, mediates the regiospecific hydrolysis of ATP to ADP when crystallized with 1 equiv of ATP.
19
Studies on a potentially prebiotic synthesis of RNA
TL;DR: A novel, potentially prebiotic synthesis of RNA is proposed in which the key steps are an aldol mode of polymerisation, intramolecular redox transfer and ring closure via a mesomeric heterocyclic betaine intermediate.
18
Patent
System for manipulating nucleic acids
Kevin A. Jarrell,Brian M. Turczyk +1 more
- 04 May 2006
TL;DR: In this paper, an improved system for linking nucleic acids to one another is presented, which provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules.
18
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