title: A screening protocol for identification of functional mutants of RNA editing adenosine deaminases.
abstract: Genetic screens can be used to evaluate a spectrum of mutations and thereby infer the function of particular residues within a protein. The Adenosine Deaminase Acting on RNA (ADAR) family of RNA-editing enzymes selectively deaminate adenosines (A) in double-helical RNA, generating inosine (I). The protocol described here exploits the editing activity of ADAR2 in a yeast-based screen by inserting an editing substrate sequence with a stop codon incorporated at the editing site upstream from the sequence encoding the reporter α-galactosidase. A-to-I editing changes the stop codon to a tryptophan codon, allowing normal expression of the reporter. This technique is particularly well-suited for screening ADAR and ADAR substrate mutant libraries for editing activity. Curr. Protoc. Chem. Biol. 4:357-369 © 2012 by John Wiley & Sons, Inc.
authors.affiliation: Department of Chemistry, University of California, Davis, Davis, California.
authors.forename: Tristan
authors.initials: T
authors.lastname: Eifler
authors.forename: Dalen
authors.initials: D
authors.lastname: Chan
authors.forename: Peter A
authors.initials: PA
authors.lastname: Beal
keywords: ADAR substrate; A‐to‐I editing; RNA editing; screen; yeast