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The CRISPR diagnostic (CRISPR-Dx) technology that employs the  trans -cleavage activities has shown great potential in diagnostic sensitivity, specificity, convenience, and portability, and has been recognized as the next-generation diagnostic methods. However, due to the lack of standardized definition of Cas  trans -cleavage enzymatic units, it is difficult to standardize the present CRISPR-Dx systems, which have undoubtedly impeded the development of the CRISPR-Dx industry. To solve the problem, we here first systematically optimized the reaction systems for Cas12a, and then defined its  trans -cleavage units ( trans U), which we believe will be of great importance and interest to researchers in both molecular diagnostic industry and basic research. Moreover, a simple protocol was provided to facilitate a step-by-step measurement of the Cas12a  trans U, which can also act as a reference for the definition of the  trans U for other Cas proteins.

Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics