<i>N</i>1-Methyladenosine detection with CRISPR-Cas13a/C2c2

Abstract

Recent studies suggested that the widespread presence of <i>N</i>1-methyladenosine (m¹A) plays a very important role in environmental stress, ribosome biogenesis and antibiotic resistance. The RNA-guided, RNA-targeting CRISPR Cas13a exhibits a "collateral effect" of promiscuous RNase activity upon target recognition with high sensitivity. Inspired by the advantage of CRISPR Cas13a, we designed a system to detect m¹A induced mismatch, providing a rapid, simple and fluorescence-based m¹A detection. For A-ssRNA, the Cas13a-based molecular detection platform showed a high fluorescence signal. For m¹A-ssRNA, there is an about 90% decline of fluorescence. Moreover, this approach can also be used to quantify m¹A in RNAs and applied for the analysis of dynamic m¹A demethylation of 28S rRNA with AlkB.