Key result
A yeast-based screening protocol exploiting ADAR2 editing activity allows for the identification of functional mutants by changing a stop codon to a tryptophan codon to express a reporter.
Describes a yeast-based screening protocol to evaluate mutations and infer the function of residues within ADAR RNA-editing enzymes.
No immediate clinical implications; extends basic tools for ADAR2 functional studies but leaves translation open.
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.
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Eifler et al. (2012) studied this question. Yeast-based screening protocol for ADAR2 editing activity was evaluated. A yeast-based screening protocol exploiting ADAR2 editing activity allows for the identification of functional mutants by changing a stop codon to a tryptophan codon to express a reporter.