Improved heat-based CRISPR method for detecting BCR1-type PML::RARα fusion and L217F mutation

A detection limit of 0.1 aM was achieved for the BCR1-type PML::RARα fusion using a novel CRISPR/Cas12a-based method.

• Moderate temperature increases improved CRISPR/Cas12a activity while enabling the use of atypical suboptimal protospacer adjacent motifs (PAMs).
• The developed method sensitively identified low-abundance mutant cells in the presence of wild-type cells.
• Efficacy was demonstrated in detecting minimal residual disease samples from patients with relapsed acute promyelocytic leukemia.
• A sensitive approach for detecting L217F single-base mutations reached a detection limit of 10 aM.
• The methods may facilitate the rapid diagnosis of acute promyelocytic leukemia and the detection of low-abundance drug-resistance mutations.

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