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Recent studies suggested that the widespread presence of N 1-methyladenosine (m 1 A) 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 m 1 A induced mismatch, providing a rapid, simple and fluorescence-based m 1 A detection. For A-ssRNA, the Cas13a-based molecular detection platform showed a high fluorescence signal. For m 1 A-ssRNA, there is an about 90% decline of fluorescence. Moreover, this approach can also be used to quantify m 1 A in RNAs and applied for the analysis of dynamic m 1 A demethylation of 28S rRNA with AlkB.

N1-Methyladenosine detection with CRISPR-Cas13a/C2c2