Kit to efficiently identify and enrich adenosine-to-inosine (A-to-I) editing in RNA molecules.
- Robust and facile method for enriching A-to-I edited transcripts.
- Overcomes the low efficiency and accuracy of transcriptome-wide RNA-seq.
A-to-I RNA editing by adenosine deaminases is a common RNA modification of double-stranded RNA. RNA editing plays critical roles in RNA metabolism, and may also be involved in cancer, aging and/or autoimmune diseases. Accurate identification and effective editing rate are necessary to understanding the effect of A-to-I RNA editing on regulation of epitranscriptome and relationships with disease. Recent developments in next generation RNA sequencing (RNA-seq) allow researchers to detect A-to-I editing. However, current RNA-seq technique is still inefficient and prone to error, underscoring the need for a simple, but robust method to enrich A-to-I edited transcripts. Currently, there are no robust methods of inosine-containing transcripts based on affinity purification.
Emory researchers have developed a method to selectively isolate A-to-I edited RNAs using Endonuclease V (eEndoV), called EndoVIPER (Endonuclease V inosine precipitation enrichment). By changing levels of Mg2+ and Ca2+, eEndoV catalysis can be switched from cleaving to promoting binding of inosine, enabling pulldown of A-to-I edited transcripts with high affinity and high specificity. They demonstrated that EndoVIPER enables isolation and enrichment of A-to-I edited transcripts from cellular RNA (brain mRNA). This method offers a robust and facile workflow for A-to-I identification and enrichment, helping to understand the regulation of A-to-I editing in a range of biological functions and disease processes.
In vitro experiments have been demonstrated with full RNA-seq validation of human brain mRNA.
Publication: Knutson, S. et al. (2019). BioRxiv.