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N1-Methyladenosine detection with CRISPR-Cas13a/C2c2

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Abstract

Recent studies suggested that the widespread presence of N1-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.

Graphical abstract: N1-Methyladenosine detection with CRISPR-Cas13a/C2c2

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Publication details

The article was received on 31 Jul 2018, accepted on 10 Jan 2019 and first published on 10 Jan 2019


Article type: Edge Article
DOI: 10.1039/C8SC03408G
Citation: Chem. Sci., 2019, Advance Article
  • Open access: Creative Commons BY-NC license
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    N1-Methyladenosine detection with CRISPR-Cas13a/C2c2

    Y. Chen, S. Yang, S. Peng, W. Li, F. Wu, Q. Yao, F. Wang, X. Weng and X. Zhou, Chem. Sci., 2019, Advance Article , DOI: 10.1039/C8SC03408G

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