Lina Li, Canxing Duan, Jianfeng Weng, Xiantao Qi, Changlin Liu, Xinhai Li, Jinjie Zhu* and Chuanxiao Xie*
SCIENCE CHINA Life Sciences, IF: 6.038
Abstract: For some Cas nucleases, trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic detection. Portable single and multiplex nucleic acid-based detection is needed forcrop pathogen management in agriculture. Here, we harnessed and characterized RfxCas13d as an additional CRISPR/Casnucleic acid detection tool. We systematically characterized AsCas12a, LbCas12a, LwaCas13a, and RfxCas13d combined withisothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen detection. Our dataindicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as input. Using this tool,we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streakeddwarf virus in crude extracts prepared in the field. Our method, from sample preparation to result readout, could be rapidly andeasily deployed in the field. This system could be extended to other crop pathogens, including those that currently lack adetection method and have metabolite profiles that make detection challenging. This nucleic acid detection system could also beused for single-nucleotide polymorphism genotyping, transgene detection, and qualitative detection of gene expression in thefield.