Article
Improving the efficiency of high-fidelity Cas9 by enhancing PAM-distal interactions.
Nature structural & molecular biology - 1 Apr 2026
Zheng Rong, Lu Zhike, Wei Rongwei, Shin Young-Cheul, Du Jiang, Zhang Qingfeng, Li Jianbo, Wang Xiaoqi, Wei Yi, Liu Botao, Chen Yang, Ding Lihong, Zhang Heng, Chen Hui, Huang Jing, Ma Lijia
Abstract excerpt
Engineering CRISPR enzymes for high fidelity often impairs cleavage activity. Meanwhile, a mechanistic understanding of why high-fidelity mutations reduce Cas9's cleavage activity remains unclear, presenting a challenge in balancing nuclease specificity and efficiency for clinical applications. In this study, we show that extending the spacer region to 21 or 22 nucleotides restores the impaired cleavage activity...
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