ICE
ICE
ICE followed by NGS identifies adenosine-to-inosine editing. In this method, RNA is treated with acrylonitrile, while control RNA is untreated. Control and treated RNAs are reverse-transcribed and PCR-amplified. Inosines in RNA fragments treated with acrylonitrile cannot be reverse-transcribed. Deep sequencing of the cDNA prepared from control and treated RNA provides high-resolution reads of inosines in RNA fragments.
Pros:
- Provides mapping of adenosine-to-inosine editing
- Can be performed with limited material
Cons:
- Nonlinear PCR amplification can lead to biases, affecting reproducibility
- Amplification errors caused by polymerases will be represented and sequenced incorrectly