Split technology in sensors based on CRISPR/Cas12a system

Jun 20, 2025Biotechnology advances

Split sensors using the CRISPR/Cas12a system

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Abstract

Split technology can enable the CRISPR/Cas12a system to detect dsDNA without PAM, short ssDNA under 15 nucleotides, and RNA.

The application of split technology enhances the stability of crRNA.
Multiple input and output logic circuits can be constructed in cells using this technology.
Cost-effective, label-free reporters can be integrated into sensors based on the CRISPR/Cas12a system.
This advancement expands the target detection range beyond the capabilities of the canonical CRISPR/Cas12a system.
Challenges and future directions for sensors utilizing this technology are discussed.

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CRISPR/Cas12a system has become a popular tool for nucleic acid analysis in recent years due to its high specificity, sensitivity and programmability. Recently, split technology has been applied to the CRISPR/Cas12a system for activators, crRNA, reporter and Cas12a. As a result, dsDNA without PAM, short ssDNA less than 15 nucleotides, and RNA can be directly detected, which are beyond the target scope of the canonical CRISPR/Cas12a system. Label-free reporter with lower cost can be incorporated into sensors based on the CRISPR/Cas12a system. Logic circuits with multiple inputs and outputs can be constructed in cells. Therefore, split technology can expand the target scope, enhance crRNA stability, increase strategy programmability, and reduce detection cost for the CRISPR/Cas12a system. In this review, we focus on the advancements of split technology in sensors based on the CRISPR/Cas12a system. We also summarize the advantages brought by split technology and discuss the challenges and perspectives of sensors based on the CRISPR/Cas12a system.

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Split technology in sensors based on CRISPR/Cas12a system

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