Leveraging Cas13a's trans-cleavage on RNA G-quadruplexes for amplification-free RNA detection

Tao Li; Dongjuan Chen; Xiaoling He; Zheyu Li; Zhichen Xu; Runchen Li; Bingxin Zheng; Rui Hu; Jiang Zhu; Ying Li; Yunhuang Yang

doi:10.1039/d3cc06238d

Leveraging Cas13a's trans-cleavage on RNA G-quadruplexes for amplification-free RNA detection

Chem Commun (Camb). 2024 Mar 14;60(23):3166-3169. doi: 10.1039/d3cc06238d.

Authors

Tao Li^{1

2}, Dongjuan Chen³, Xiaoling He^{1

2}, Zheyu Li^{1

2}, Zhichen Xu^{1

2}, Runchen Li^{1

2}, Bingxin Zheng^{1

2}, Rui Hu^{1

2}, Jiang Zhu^{1

2}, Ying Li^{1

2}, Yunhuang Yang^{1

2

4}

Affiliations

¹ State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology-Wuhan National Laboratory for Optoelectronics, Chinese Academy of Sciences, Wuhan 430071, China. liying@wipm.ac.cn.
² University of Chinese Academy of Sciences, Beijing 100049, China.
³ Department of Laboratory Medicine, Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430070, China.
⁴ Optics Valley Laboratory, Hubei 430074, China.

PMID: 38410041
DOI: 10.1039/d3cc06238d

Abstract

In this study, we investigated Cas13a's efficacy in trans-cleaving RNA G-quadruplexes (rG4s) as an alternative to ssRNA reporters in CRISPR-Cas13a diagnostics. Our findings demonstrate enhanced efficiency due to the structural arrangement of rG4s. Implementing a simplified CRISPR-Cas13a system based on rG4, we identified SARS-CoV-2 infections in 25 patient samples within 1 hour without target pre-amplification.

MeSH terms

COVID-19* / diagnosis
Clustered Regularly Interspaced Short Palindromic Repeats
G-Quadruplexes*
Humans
RNA / chemistry
RNA / genetics

Substances

RNA