Evolution of a General RNA-Cleaving FANA Enzyme.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 30498223.
- Also identified by DOI 10.1038/s41467-018-07611-1 and PMC identifier 6265334.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
The isolation of synthetic genetic polymers (XNAs) with catalytic activity demonstrates that catalysis is not limited to natural biopolymers, but it remains unknown whether such systems can achieve robust catalysis with Michaelis-Menten kinetics. Here, we describe an efficient RNA-cleaving 2'-fluoroarabino nucleic acid enzyme (FANAzyme) that functions with a rate enhancement of >10<sup>6</sup>-fold over the uncatalyzed reaction and exhibits substrate saturation kinetics typical of most natural enzymes. The FANAzyme was generated by in vitro evolution using natural polymerases that were found to recognize FANA substrates with high fidelity. The enzyme comprises a small 25 nucleotide catalytic domain flanked by substrate-binding arms that can be engineered to recognize diverse RNA targets. Substrate cleavage occurs at a specific phosphodiester bond located between an unpaired guanine and a paired uracil in the substrate recognition arm. Our results expand the chemical space of nucleic acid enzymes to include nuclease-resistant scaffolds with strong catalytic activity.
Medical subject headings
- Nucleic Acids
- RNA