CRISPR/Cas systems accelerating the development of aptasensors

https://doi.org/10.1016/j.trac.2022.116775Get rights and content

Highlights

  • The review summarizes the recent progress of CRISPR/Cas system-based aptasensors including their design principles and superior applications.

  • The review emphasizes eight types of Cas-aptasensor on their respects of fabrication methods, bio-recognition mechanism, as well as detection evaluation.

  • The challenges and future directions are concluded and depicted for more applications of Cas-aptasensors.

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (CRISPR/Cas) systems, regarded as “God's scissors”, have recently received considerable attention in performing genome editing, transcriptional regulation and biosensor construction, benefiting from their adjustable mechanical properties, easy to operate and design, good biocompatibility, and the collateral cleavage activity. Since 2019, aptamers have been becoming an attractive star integrated into CRISPR/Cas systems as newly emerging molecular recognition elements and flexible signal transduction modules, with their significant advantages such as high sensitivity, remarkable specificity, in vitro synthesis, base-pairing, labeling and modification, and programmability capability. In this review, the recent progress of CRISPR/Cas system-based aptasensors (Cas-aptasensors) is comprehensively summarized including their design principles and superior applications. Firstly, we briefly introduce the essential features of aptamers and the CRISPR/Cas system and then outline the composition of Cas-aptasensors involving the Cas proteins, crRNA, reporter probes, as well as the target analytes and their specific aptamers. In detail, we emphasize their fabrication methods, bio-recognition mechanism, as well as the detection evaluation of four major Cas-aptasensors types of fluorescent, electrochemical, colorimetric, and upconversion luminescent resonance energy transfer, and other four novel Cas-aptasensors including inductively light-up RNA aptamer-based sensors coupled plasma mass spectrometry, resonance Rayleigh scattering, and surface-enhanced Raman scattering, along with. Finally, the challenges and future directions are concluded and depicted for aptamers, CRISPR/Cas systems, and their accelerating applications in Cas-aptasensors. This review is expected to inspire more researchers to have insight into Cas-aptasensors and to help to improve the bioanalytical efficiency and probability of success.

Section snippets

Introduction of CRISPR/Cas and aptamer

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated nucleases proteins, termed as CRISPR/Cas systems [1,2], are the RNA-guided adaptive immunity mechanism formed in the long-term evolution of archaea and bacteria. Since its serendipitous discovery by Nakata's group in 1987 [1], scientists have been studying its function and mechanism and have gradually gained a relatively comprehensive understanding [[3], [4], [5]]. In August 2012, Jennifer Doudna and

The essential composition of Cas-aptasensors

CRISPR/Cas system is composed of palindromic sequences regions, spacer regions (exogenous DNA sequences), the upstream leader region corresponding to the promoter of the CRISPR sequence, a polymorphic family of genes (Cas genes) and Cas proteins [[5], [6], [7]]. Based on the different functions of Cas proteins and the mechanism of action, CRISPR/Cas system can be broadly classified into two categories: the Class 1 is a multi-protein effector system requiring 4–7 Cas protein subunits considered

CRISPR/Cas system-based aptasensors

CRISPR/Cas system has been rising since 2019 as the attractive stars in aptamers development with their superior properties. Based upon their methods of signal transduction, we classify these aptasensors into four major types of fluorescent, electrochemical, colorimetric, LERT, and other three types involving ICPMS, RRS, and SERS sensors. Besides, these Cas-aptasensors present many advantages such as rapid, sensitive, tunable, versatility, liquid biopsy and low cost, and provide miniaturized

Conclusion and future perspectives

To sum up, this review has summarized the recent advances in CRISPR/Cas system-enhanced aptasensors for the detection of a broad range of target analytes. Unique characteristics of CRISPR/Cas system have made them ideal for aptasensors fabrication, biological recognition and signal transduction. They have largely improved the sensitivity in bioanalysis coupled with some flexible technologies for yielding more acDNA, such as “locked-activated” mode, sandwich-type CLASA, “core-shell” nanoprobe

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

All authors made the contributions to the manuscript and had approved the final version of the script. We thank the support of the Natural Science Foundation of Shandong Province (no. ZR2022QB207), National Natural Science Foundation of China (no. 321020882187401021827810), Supported by the Taishan Scholars Program, the Agricultural Scientific and Technological Innovation Project of Shandong Academy of Agricultural Sciences (CXGC2021B14, CXGC2022E05), the Young Elite Scientist Sponsorship

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