Ruiwei Hu, Wei Yang, Jia Li, Lanxin Jiang, Menghan Li, Mengxuan Zhang, Yuexi Kang, Xiaoxue Cheng, Shasha Zhu, Lina Zhao, Wen He, Minghui Guo, Shijia Ding, Haiping Wu, Wei Cheng
Existing RNA in situ imaging strategies mostly utilize parallel repetitive nucleic acid self-assembly to achieve multiple analysis, with limitations of complicated systems and cumbersome steps. Here, a Cas9 code key system with key probe (KP) encoder and CRISPR/Cas9 signal exporter is developed. This system triggers T-protospacer adjacent motif (T-PAM structural transitions of multiple KP encoders to form coding products with uniform single-guide RNA (sgRNA) target sequences as tandem nodes. Only single sgRNA/Cas9 complex is required to cleave multiple coding products, enabling efficient "many-to-one" tandem signaling, and non-collateral cleavage activity-dependent automatic signaling output through active introduction of mismatched bases...
May 3, 2024: Small Methods