ICE

ICE

Inosine chemical erasing (ICE)  identifies adenosine to inosine editing. In this method, RNA is treated with acrylonitrile, while control RNA is untreated. Control and treated RNAs are then reverse-transcribed and PCR-amplified. Inosines in RNA fragments treated with acrylonitrile cannot be reverse-transcribed. Deep sequencing of the cDNA of control and treated RNA provides high-resolution reads of inosines in RNA fragments.

Pros:

• Mapping of adenosine to inosine editing
• Can be performed with limited material

Cons:

• Non-linear PCR amplification can lead to biases, affecting reproducibility
• Amplification errors caused by polymerases will be represented and sequenced incorrectly

• ICE: Sakurai M., Yano T., Kawabata H., Ueda H. and Suzuki T. (2010) Inosine cyanoethylation identifies A-to-I RNA editing sites in the human transcriptome. Nat Chem Biol 6: 733-740

1. Suzuki T., Ueda H., Okada S. and Sakurai M. (2015) Transcriptome-wide identification of adenosine-to-inosine editing using the ICE-seq method. Nat Protoc 10: 715-732
2. Sakurai M., Ueda H., Yano T., Okada S., Terajima H., et al. (2014) A biochemical landscape of A-to-I RNA editing in the human brain transcriptome. Genome Res 24: 522-534