文献类型： 外文期刊
作者： Xiaohan Tong;Tianle Li;Kun Zhang;Dongming Zhao;Ying Zhang;Hao Yin
作者机构：
期刊名称： Cell Insight
ISSN： 2772-8927
年卷期： 2025 年 4 卷 2 期
页码：
收录情况： 无来源刊（2025版）
摘要： CRISPR-Cas12a holds promising potential for pathogen detection. However, its performance is not optimal when combined with isothermal amplification. Hence, we engineered a mutant of LbCas12a (K595A) with reduced cis-cleavage activity, to minimize interference with isothermal amplification. Compared to wild-type Cas12a, the K595A mutant exhibited a 2–3 times faster reaction speed and a 1,000–10,000 times increase in sensitivity in a one-pot reaction. We applied this mutant for detection of African Swine Fever Virus (ASFV). This K595A mutant successfully detected all 30 ASFV samples within 20 minutes. Our study suggests a universal approach to improve the performance of Cas12a for pathogen detection.
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