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RNA and DNA Editing

Volume 718 of the series Methods in Molecular Biology pp 41-73

Date:

Perturbing A-to-I RNA Editing Using Genetics and Homologous Recombination

  • Cynthia J. StaberAffiliated withMolecular Biology Cellular Biology and Biochemistry, Brown University
  • , Selena GellAffiliated withMolecular Biology Cellular Biology and Biochemistry, Brown University
  • , James E. C. JepsonAffiliated withMolecular Biology Cellular Biology and Biochemistry, Brown University
  • , Robert A. ReenanAffiliated withMolecular Biology Cellular Biology and Biochemistry, Brown University Email author 

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Abstract

Evidence for the chemical conversion of adenosine-to-inosine (A-to-I) in messenger RNA (mRNA) has been detected in numerous metazoans, especially those “most successful” phyla: Arthropoda, Mollusca, and Chordata. The requisite enzymes for A-to-I editing, ADARs (adenosine deaminases acting on RNA) are highly conserved and are present in every higher metazoan genome sequenced to date. The fruit fly, Drosophila melanogaster, represents an ideal model organism for studying A-to-I editing, both in terms of fundamental biochemistry and in relation to determining adaptive downstream effects on physiology and behavior. The Drosophila genome contains a single structural gene for ADAR (dAdar), yet the fruit fly transcriptome has the widest range of conserved and validated ADAR targets in coding mRNAs of any known organism. In addition, many of the genes targeted by dADAR have been genetically identified as playing a role in nervous system function, providing a rich source of material to investigate the biological relevance of this intriguing process. Here, we discuss how recent advances in the use of ends-out homologous recombination (HR) in Drosophila make possible both the precise control of the editing status for defined adenosine residues and the engineering of flies with globally altered RNA editing of the fly transcriptome. These new approaches promise to significantly improve our understanding of how mRNA modification contributes to insect physiology and ethology.

Key words

Drosophila melanogaster RNA editing Homologous recombination