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

Chao-Qun Yuan, Tian Tian, Jian Sun, Meng-Lu Hu, Xu-Sheng Wang, Er-Hu Xiong, Meng Cheng, Yi-Juan Bao, Wei Lin, Jie-Ming Jiang, Cheng-Wei Yang, Qian Chen (+7 others)
2019 bioRxiv   pre-print
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
more » ... equence 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.
doi:10.1101/615724 fatcat:hqd2quj7ljdedmfd47peiq4li4