Isolation of new ribozymes from a large pool of random sequences [see comment].

  title={Isolation of new ribozymes from a large pool of random sequences [see comment].},
  author={David P. Bartel and Jack W. Szostak},
  volume={261 5127},
An iterative in vitro selection procedure was used to isolate a new class of catalytic RNAs (ribozymes) from a large pool of random-sequence RNA molecules. These ribozymes ligate two RNA molecules that are aligned on a template by catalyzing the attack of a 3'-hydroxyl on an adjacent 5'-triphosphate--a reaction similar to that employed by the familiar protein enzymes that synthesize RNA. The corresponding uncatalyzed reaction also yields a 3',5'-phosphodiester bond. In vitro evolution of the… 

A ribozyme that ligates RNA to protein

  • S. BaskervilleD. Bartel
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 2002
An optimized version of the ribozyme reacts with substrate-fusion proteins, allowing it to be used to attach RNA tags to proteins both in vitro and within bacterial cells, suggesting a simple way to tag a specific protein with amplifiable information.

In vitro evolution of new ribozymes with polynucleotide kinase activity

An engineered version of one class of kinases is able to catalyse the transfer of thiophosphate from ATP-γS to the 5′-hydroxyl of an exogenous oligoribonucleotide substrate with multiple turnover, thus acting as a true enzyme.

Creation and evolution of new ribozymes.

An RNA is generated that synthesizes RNA using the same reaction as that employed by protein enzymes that catalyze RNA polymerization, and shows significant template fidelity.

In vitro evolution of a self-alkylatlng ribozyme

RNA enzymes are postulated to have catalysed all chemical reactions in the earliest living cells, and selection for self-biotinylation yields a transfer RNA-like ribozyme that efficiently catalyses carbon–nitrogen bond formation.

The New World of ribozymes.

  • L. Jaeger
  • Biology
    Current opinion in structural biology
  • 1997

Ribozyme-Catalyzed Transcription of an Active Ribozyme

The evolution and engineering of an RNA polymerase ribozyme capable of synthesizing RNAs of up to 95 nucleotides in length is described, and the accurate synthesis of an enzymatically active RNA, a hammerhead endonuclease ribo enzyme is demonstrated.

Ribozyme-catalysed amino-acid transfer reactions

In vitro selection and evolution is used to isolate ribozymes with acyl transferase activity from a pool of random RNA sequences and one of them transfers an amino acid to itself in a reaction that is analogous to peptidyl transfer on the ribosome.

New ligase-derived RNA polymerase ribozymes.

Eight ligase-derived polymerase ribozymes isolated from a pool of variants of an existing RNA ligase ribozyme are reported, each a new potential starting point for further in vitro evolution and engineering and together substantially enrich the set of candidates from which an RNA replicaseribozyme might eventually emerge.

Conversion of a ribozyme to a deoxyribozyme through in vitro evolution.




Directed evolution of an RNA enzyme.

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.

RNA as an RNA polymerase: net elongation of an RNA primer catalyzed by the Tetrahymena ribozyme.

The demonstration that an RNA enzyme can catalyze net elongation of an RNA primer supports theories of prebiotic RNA self-replication.

A multisubunit ribozyme that is a catalyst of and template for complementary strand RNA synthesis.

Results suggest that prebiotically synthesized oligonucleotides might have been able to assemble into a complex capable of self-replication.

Selection of a ribozyme that functions as a superior template in a self-copying reaction.

In vitro genetic selection was used to identify mutations that reactivate a virtually inactive sunY deletion mutant and a selected mutant with five substitution mutations scattered throughout the primary sequence showed greater catalytic activity than the original ribozyme under the selection conditions.

In vitro selection of RNAs that undergo autolytic cleavage with Pb2+.

Both specific mutations and terminal truncation experiments suggest that the D and T loops of these two variants interact in a manner similar to that of tRNA(Phe) despite the absence of the G18U55 and G19C56 tertiary interactions.

Catalysis of RNA cleavage by the Tetrahymena thermophila ribozyme. 1. Kinetic description of the reaction of an RNA substrate complementary to the active site.

Mechanistic data suggest that binding of the oligonucleotide substrate, G2CCCUCUA5, and binding of G are essentially random and independent, and tertiary interactions are involved in binding.

RNA catalysis and the origins of life.

  • L. Orgel
  • Biology, Chemistry
    Journal of theoretical biology
  • 1986

Evolution in vitro of an RNA enzyme with altered metal dependence

Variants of the Tetrahymena ribozyme are obtained that are capable of cleaving an RNA substrate in reaction mixtures containing Ca2+ as the divalent cation, extending the range of different chemical environments available to RNA enzymes.

A small metalloribozyme with a two-step mechanism

An RNA molecule consisting of an asymmetric internal loop of six nucleotides can be rapidly and specifically cleaved by Pb2+ in the presence of Mg2+. The 5′ cleavage product terminates with a 3′

Activation of the catalytic core of a group I intron by a remote 3' splice junction.

It is proposed that this novel interaction is necessary to drive 5' exon-core transcripts into an active conformation and a requirement for additional stabilizing interactions appears to be a general feature of group I self-splicing.