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CoHIT: a one-pot ultrasensitive ERA-CRISPR system for detecting multiple same-site indels

Nat Commun. 2024-06; 
Yin Liu, Xinyi Liu, Dongyi Wei, Lu Dang, Xiaoran Xu, Shisheng Huang, Liwen Li, Sanyun Wu, Jinxian Wu, Xiaoyan Liu, Wenjun Sun, Wanyu Tao, Yongchang Wei, Xingxu Huang, Kui Li, Xinjie Wang, Fuling Zhou
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Gene Synthesis … small indel detection were directly synthetized by GenScript (Nanjing, China), and the sequences are listed in Supplementary Data 5. The plasmids carrying the NPM1 gene WT and c.… Get A Quote

摘要

Genetic testing is crucial for precision cancer medicine. However, detecting multiple same-site insertions or deletions (indels) is challenging. Here, we introduce CoHIT (Cas12a-based One-for-all High-speed Isothermal Test), a one-pot CRISPR-based assay for indel detection. Leveraging an engineered AsCas12a protein variant with high mismatch tolerance and broad PAM scope, CoHIT can use a single crRNA to detect multiple NPM1 gene c.863_864 4-bp insertions in acute myeloid leukemia (AML). After optimizing multiple parameters, CoHIT achieves a detection limit of 0.01% and rapid results within 30?minutes, without wild-type cross-reactivity. It successfully identifies NPM1 mutations in 30 out of 108 AML patients a... More

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