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Article

Iterative crRNA design and a PAM-free strategy enabled an ultra-specific RPA-CRISPR/Cas12a detection platform.

Communications biology - 7 Nov 2024

Mao Xujian, Xu Jian, Jiang Jingyi, Li Qiong, Yao Ping, Jiang Jinyi, Gong Li, Dong Yin, Tu Bowen, Wang Rong, Tang Hongbing, Yao Fang, Wang Fengming

Abstract excerpt

CRISPR/Cas12a is a highly promising detection tool. However, detecting single nucleotide variations (SNVs) remains challenging. Here, we elucidate Cas12a specificity through crRNA engineering and profiling of single- and double-base mismatch tolerance across three targets. Our findings indicate that Cas12a specificity depends on the number, type, location, and distance of mismatches within the R-loop. We also...

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Identifiers

PMID: 39506042
DOI: 10.1038/s42003-024-07173-7
PMCID: PMC11541961

Genes

PAM
GC

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