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bioRxiv · 10.1101/615724

Universal and naked-eye gene detection platform based on CRISPR/Cas12a/13a system

Yuan, C.-Q.·Tian, T.·Sun, J.·Hu, M.-L.·Wang, X.-S.·Xiong, E.-H.·Cheng, M.·Bao, Y.-J.·Lin, W.·Jiang, J.-M.·Yang, C.-W.·Chen, Q.·Wang, H.·Wang, X.-R.·Deng, X.-B.·Liao, X.-P.·Liu, Y.-H.·Zhang, G.-H.·Zhou, X.
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Abstract

Colorimetric gene detection based on gold nanoparticles (AuNPs) is an attractive detection format due to its simplicity. Here, we report a new design for a colorimetric gene-sensing platform based on the CRISPR/Cas system that has improved specificity, sensitivity, and universality. CRISPR/Cas12a and CRISPR/Cas13a have two distinct catalytic activities and are used for specific target gene recognition. Programmable recognition of DNA by Cas12a/crRNA and RNA by Cas13a/crRNA with a complementary sequence activates the nonspecific trans-ssDNA or -RNA cleavage, respectively, thus degrading the ssDNA or RNA linkers which are designed as a hybridization template for the AuNP-DNA probe pair. Target-induced trans -ssDNA or RNA cleavage leads to a distance-dependent color change for the AuNP-DNA probe pair. In this platform, naked eye detection of transgenic rice, African swine fever virus (ASFV), and a miRNA can be completed within 1 hour. Our colorimetric gene-sensing method shows superior characteristics, such as probe universality, isothermal reaction conditions, on-site detection capability, and sensitivity that is comparable to that of the fluorescent detection; thus, this method represents a robust next generation gene detection platform.

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Yuan, C.-Q., Tian, T., Sun, J., Hu, M.-L., Wang, X.-S., Xiong, E.-H., Cheng, M., Bao, Y.-J., Lin, W., Jiang, J.-M., Yang, C.-W., Chen, Q., Wang, H., Wang, X.-R., Deng, X.-B., Liao, X.-P., Liu, Y.-H., Zhang, G.-H., Zhou, X.. 2019-05-07. Universal and naked-eye gene detection platform based on CRISPR/Cas12a/13a system. https://doi.org/10.1101/615724

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