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
Ribozymes with RNA protein binding site
John M. Burke,Bruno Sargueil +1 more
- 27 Dec 1995
TL;DR: Ribozyme having a ligand binding site formed as a doublestranded RNA and a single-stranded loop, the ribozyme having enzymatic activity to cleave and/or ligate itself or a separate RNA molecule as mentioned in this paper.
34
The RNA-world and co-evolution hypotheses and the origin of life: implications, research strategies and perspectives.
TL;DR: A central element in the proposed experimental strategies is the study of the catalytic activities of very small peptides and RNA-like oligomers, according to existing, as well as to yet-to-be-invented scenarios of the two hypotheses under consideration.
33
Titratable Avidity Reduction Enhances Affinity Discrimination in Mammalian Cellular Selections of Yeast-Displayed Ligands.
Lawrence A. Stern,Clifford M. Csizmar,Daniel R. Woldring,Carston R. Wagner,Benjamin J. Hackel +4 more
TL;DR: A facile approach for affinity selection of yeast-displayed ligands against full-length cellular targets is offered and the effectiveness of this method is demonstrated by generating EpCAM-binding ligands that are promising for further applications.
33
On the levels of enzymatic substrate specificity: Implications for the early evolution of metabolic pathways
TL;DR: It is argued that both sequence comparisons and experimental results on enzyme substrate specificity support the patchwork assembly theory, which suggests that gene duplication events followed by a gradual neoDarwinian accumulation of mutations and other minute genetic changes lead to the narrowing and modification of enzyme function in at least some primordial metabolic pathways.
32
Nanofabrication with biomolecules
Dominic C. Chow,Matthew S. Johannes,Woo-Kyung Lee,Robert L. Clark,Stefan Zauscher,Ashutosh Chilkoti +5 more
TL;DR: To provide useful insights into the future directions of bionanofabrication, current developments in characterization techniques, fabrication methods, and their integration and control in manufacturing processes are discussed.
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