Abstract
The conversion of adenosine to inosine (A to I) by RNA editing represents a common posttranscriptional mechanism for diversification of both the transcriptome and proteome, and is a part of the cellular response for innate immune tolerance. Due to its preferential base-pairing with cytosine (C), inosine (I) is recognized as guanosine (G) by reverse transcriptase, as well as the cellular splicing and translation machinery. A-to-I editing events appear as A-G discrepancies between genomic DNA and cDNA sequences. Molecular analyses of RNA editing have leveraged these nucleoside differences to quantify RNA editing in ensemble populations of RNA transcripts and within individual cDNAs using high-throughput sequencing or Sanger sequencing-derived analysis of electropherogram peak heights. Here, we briefly review and compare these methods of RNA editing quantification, as well as provide experimental protocols by which such analyses may be achieved.
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Acknowledgments
This work was supported by the Joel G. Hardman and Mary K. Parr Endowed Chair in Pharmacology, NIH/NIDDK (R01 DK119508), and Vanderbilt Technologies for Advanced Genomics [VANTAGE, Vanderbilt Ingram Cancer Center (P30 CA68485), the Vanderbilt Vision Center (P30 EY08126), and NIH/NCRR (G20 RR030956)].
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Malik, T.N., Cartailler, JP., Emeson, R.B. (2021). Quantitative Analysis of Adenosine-to-Inosine RNA Editing. In: Picardi, E., Pesole, G. (eds) RNA Editing. Methods in Molecular Biology, vol 2181. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0787-9_7
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DOI: https://doi.org/10.1007/978-1-0716-0787-9_7
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