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
Self-Organizing Algorithms Derived from RNA Interactions
Wolfgang Banzhaf
- 01 Jan 1995
TL;DR: This work discusses algorithms based on the RNA interaction found in Nature and introduces folding methods to achieve 2-dimensional alternative forms of the sequences and follows the evolution of a number of sample simulations and analyse the resulting self-organising system.
18
Patent
Methods and reagents for oligonucleotide synthesis
Chandra Vargeese,Weimin Wang +1 more
- 03 Jul 2002
TL;DR: In this paper, the authors present novel compositions, linkers, derivatized solid supports, and methods for the efficient solid phase synthesis of oligonucleotides, including RNA, DNA, RNA-DNA chimeras, and analogs thereof.
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Combinatorial Chemistry in Biology
Michael Famulok,Ernst-Ludwig Winnacker,Chi-Huey Wong +2 more
- 01 Jan 1999
TL;DR: The Two Hybrid Toolbox and Evolutionary Approaches to Protein Engineering are reviewed.
In Vitro Selection of Nucleic Acid Enzymes
Kurz M,Ronald R. Breaker +1 more
TL;DR: Over the course of 4 billion years of evolution, it appears that nature has determined that proteins are a superior format for constructing enzymes; however, the true capacity of nucleic acids for catalytic function remains to be fully defined.
18
Evolution of an Enzyme from a Noncatalytic Nucleic Acid Sequence.
TL;DR: The findings demonstrate that an enzyme can indeed arise from a sequence of a functional polymer via permissive molecular evolution, a mechanism that may have been exploited by nature for the creation of the enormous repertoire of enzymes in the biological world today.
References
DNA sequencing with chain-terminating inhibitors
TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase
Craig Tuerk,Larry Gold +1 more
TL;DR: High-affinity nucleic acid ligands for a protein were isolated by a procedure that depends on alternate cycles of ligand selection from pools of variant sequences and amplification of the bound species.
10.4K
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
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TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
9.7K
In vitro selection of RNA molecules that bind specific ligands.
TL;DR: Subpopulations of RNA molecules that bind specifically to a variety of organic dyes have been isolated from a population of random sequence RNA molecules.
9.7K