PUBLICATION
A real-time PCR method for the quantitative analysis of RNA editing at specific sites
- Authors
- Chen, Y.C., Kao, S.C., Chou, H.C., Lin, W.H., Wong, F.H., and Chow, W.Y.
- ID
- ZDB-PUB-080227-9
- Date
- 2008
- Source
- Analytical biochemistry 375(1): 46-52 (Journal)
- Registered Authors
- Keywords
- Quantitative PCR, RNA editing, Zebrafish, Ionotropic glutamate receptor, Real-time PCR
- MeSH Terms
-
- Animals
- Base Sequence
- DNA Primers/metabolism
- Gene Expression Regulation, Developmental
- Molecular Sequence Data
- Polymerase Chain Reaction/methods*
- RNA/analysis*
- RNA Editing/genetics*
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- Receptors, Glutamate/genetics
- Receptors, Glutamate/metabolism
- Reproducibility of Results
- Zebrafish/embryology
- Zebrafish/genetics
- PubMed
- 18252192 Full text @ Anal. Biochem.
Citation
Chen, Y.C., Kao, S.C., Chou, H.C., Lin, W.H., Wong, F.H., and Chow, W.Y. (2008) A real-time PCR method for the quantitative analysis of RNA editing at specific sites. Analytical biochemistry. 375(1):46-52.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping