title: A real-time PCR method for the quantitative analysis of RNA editing at specific sites.
abstract: In this study, a quantitative PCR (qPCR) method was developed to determine the A-to-I RNA editing frequencies at specific sites. The A-to-I RNA editing of nuclear transcripts exerts profound effects on the biological activities of gene products. RNA editing of nuclear gene transcripts have been shown to be developmentally regulated and tissue specific, and alternations of RNA editing activities have been observed under pathological conditions. Two sites of ionotropic glutamate receptor subunits, the Q/R site of zebrafish gria2alpha and the Y/C site of grik2alpha, were chosen in this study to demonstrate the applicability of the SYBR Green detection-based real-time PCR method to measure RNA editing activities during zebrafish development. The results obtained by qPCR were consistent with those obtained by the limited primer extension. However, the qPCR method has the advantages of easy handling and low cost.
authors.affiliation: Department of Life Sciences, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan.
authors.forename: Yu-Chia
authors.initials: YC
authors.lastname: Chen
authors.forename: Shang-Ching
authors.initials: SC
authors.lastname: Kao
authors.forename: Hsiao-Chin
authors.initials: HC
authors.lastname: Chou
authors.forename: Wei-Hsiang
authors.initials: WH
authors.lastname: Lin
authors.forename: Feng-Hua
authors.initials: FH
authors.lastname: Wong
authors.forename: Wei-Yuan
authors.initials: WY
authors.lastname: Chow
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000032:analysis*; D017393:RNA Editing / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017470:Receptors, Glutamate / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D015027:Zebrafish / Q000196:embryology / Q000235:genetics
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D017470:Receptors, Glutamate; D012313:RNA