title: A method to identify RNA A-to-I editing targets using I-specific cleavage and exon array analysis.
abstract: RNA A-to-I editing is the most common single-base editing in the animal kingdom. Dysregulations of RNA A-to-I editing are associated with developmental defects in mouse and human diseases. Mouse knockout models deficient in ADAR activities show lethal phenotypes associated with defects in nervous system, failure of hematopoiesis and reduced tolerance to stress. While several methods of identifying RNA A-to-I editing sites are currently available, most of the critical editing targets responsible for the important biological functions of ADARs remain unknown. Here we report a method to systematically analyze RNA A-to-I editing targets by combining I-specific cleavage and exon array analysis. Our results show that I-specific cleavage on editing sites causes more than twofold signal reductions in edited exons of known targets such as Gria2, Htr2c, Gabra3 and Cyfip2 in mice. This method provides an experimental approach for genome-wide analysis of RNA A-to-I editing targets with exon-level resolution. We believe this method will help expedite inquiry into the roles of RNA A-to-I editing in various biological processes and diseases.
authors.affiliation: Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung City 807, Taiwan. cntseng@kmu.edu.tw
authors.forename: Chao-Neng
authors.initials: CN
authors.lastname: Tseng
authors.forename: Hsueh-Wei
authors.initials: HW
authors.lastname: Chang
authors.forename: Joel
authors.initials: J
authors.lastname: Stocker
authors.forename: Hui-Chun
authors.initials: HC
authors.lastname: Wang
authors.forename: Chiu-Chin
authors.initials: CC
authors.lastname: Lu
authors.forename: Cheng-Hsuan
authors.initials: CH
authors.lastname: Wu
authors.forename: Jyuer-Ger
authors.initials: JG
authors.lastname: Yang
authors.forename: Chung-Lung
authors.initials: CL
authors.lastname: Cho
authors.forename: Hurng-Wern
authors.initials: HW
authors.lastname: Huang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001921:Brain / Q000166:cytology; D002460:Cell Line; D005091:Exons; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009419:Nerve Tissue Proteins / Q000235:genetics*; D059367:RNA Cleavage; D017393:RNA Editing; D018091:Receptors, AMPA / Q000235:genetics*; D011963:Receptors, GABA-A / Q000235:genetics*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C529497:Cyfip2 protein, mouse; C546258:Gabra3 protein, mouse; D009419:Nerve Tissue Proteins; D018091:Receptors, AMPA; D011963:Receptors, GABA-A; C104722:glutamate receptor ionotropic, AMPA 2