Frontiers in Microbiology (Nov 2021)

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

  • Hailong Lv,
  • Hailong Lv,
  • Jian Wang,
  • Jian Zhang,
  • Yijian Chen,
  • Yijian Chen,
  • Lei Yin,
  • Dian Jin,
  • Dayong Gu,
  • Huailong Zhao,
  • Yong Xu,
  • Jin Wang,
  • Jin Wang

DOI
https://doi.org/10.3389/fmicb.2021.766464
Journal volume & issue
Vol. 12

Abstract

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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 (transU), 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 transU, which can also act as a reference for the definition of the transU for other Cas proteins.

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