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Engineering CRISPR guide RNAs for programmable RNA sensors.

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As the most valuable feature of the CRISPR system, the programmability based on Watson-Crick base pairing has been widely exploited in engineering RNA sensors. The base pairing in these systems offers a connection between the RNA of interest and the CRISPR effector, providing a highly specific mechanism for RNA detection both in vivo and in vitro. In the last decade, despite the many successful RNA sensing approaches developed during the era of CRISPR explosion, a deeper understanding of the characteristics of CRISPR systems and the continuous expansion of the CRISPR family members indicates that the CRISPR-based RNA sensor remains a promising area from which a variety of new functions and applications can be engineered. Here, we present a systematic overview of the various strategies of engineering CRISPR gRNA for programmable RNA detection with an aim to clarify the role of gRNA's programmability among the present limitations and future development of CRISPR-enabled RNA sensors.


Peer reviewed: True

Acknowledgements: For the purpose of open access, the MRC Laboratory of Molecular Biology has applied a CC BY public copyright license to any Author Accepted Manuscript version arising. This work was supported as a research collaboration between AstraZeneca UK Limited and the Medical Research Council-MRC-AstraZeneca Blue Sky Grant (Y.L. and W.L.). B.W. acknowledges support by the Natural Science Foundation of China (32271475), the Fundamental Research Funds for the Central Universities (226-2022-00178, 226-2022-00214) and the Kunpeng Action Program Award of Zhejiang Province.

Publication status: Published


CRISPR, RNA sensor, gRNA, programmability, Base Pairing, CRISPR-Cas Systems, RNA, RNA, Guide, CRISPR-Cas Systems

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Biochem Soc Trans

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Portland Press Ltd.