Biomimetic Chip Enhanced Time-Gated Luminescent CRISPR-Cas12a Biosensors under Functional DNA Regulation

Abstract

Despite that the currently discovered CRISPR-Cas12a system is beneficial for improving the detection accuracy and design flexibility of luminescent biosensors, there are still challenges to extend target species and strengthen adaptability in complicated biological media. To conquer these obstacles, we present here some useful strategies. For the former, the limitation to nucleic acids assay is broken through by introducing a simple functional DNA regulation pathway to activate the unique trans-cleavage effect of this CRISPR system, under which the expected biosensors are capable of effectively transducing a protein (employing dual aptamers) and a metal ion (employing DNAzyme). For the latter, a time-gated luminescence resonance energy transfer imaging manner using a long-persistent nanophosphor as the energy donor is performed to completely eliminate the background interference and a nature-inspired biomimetic periodic chip constructed by photonic crystals is further combined to enhance the persistent luminescence. In line with the above efforts, the improved CRISPR-Cas12a luminescent biosensor not only exhibits a sound analysis performance toward the model targets (carcinoembryonic antigen and Na+) but also owns a strong anti-interference feature to actualize accurate sensing in human plasma samples, offering a new and applicative analytical tool for laboratory medicine.

A one-tube dual-readout biosensor for detection of nucleic acids and non-nucleic acids using CRISPR-ALP tandem assay

“…[9][10][11] In this regard, various CRISPR-based diagnostic platforms, including colorimetry, electrochemistry, electrochemiluminescence, and chemiluminescence, have been proposed. [12][13][14][15][16][17][18][19][20][21] Although the aforementioned biosensors can provide highly sensitive, selective, fast, and convenient testing, they are only available in a single-readout mode that is not conducive to the accuracy of detection due to the influence of environmental and instrumental factors. There is an urgent demand for extending and building a novel CRISPR/Cas12abased multiple-readout biosensor.…”

CRISPR‐Cas system manipulating nanoparticles signal transduction for cancer diagnosis

“…The detection of emerging cancer biomarkers (Hartwell et al, 2006; Hayes et al, 2014; Henry & Hayes, 2012); such as proteins (Sardesai et al, 2011; Tang et al, 2016; Wang et al, 2011), nucleic acids (Choi, Ha, et al, 2021; Choi, Lim, et al, 2021; D. Zhang et al, 2020; G. Zhang et al, 2020), and extracellular vesicles (Cleris et al, 2019; Li, Zheng, Li, et al, 2021; Li, Zheng, Lu, et al, 2021; Li, Xing, Xu, et al, 2021) provides new opportunities for early diagnosis and intervention and insights into the ongoing pathological processes. To this end, various biosensors with different signal transducers have been designed for non‐invasive and direct tracing biomolecules or molecular events in cancer cells and relevant tissues.…”

Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins

“…20 The CRISPR/Cas12a system is based on the Cas12a enzyme to indiscriminately cleave surrounding single-stranded DNA (ssDNA) probes when the functional DNA encounters the target analyte. 21,22 Furthermore, Cas12a has low intrinsic tolerance for crRNA-target DNA mismatches and requires high complementarity, which makes the Cas12a-based detection method have high specificity. 23 In addition, the sensing is not constrained on the interface space, thereby significantly improving the recognition efficiency.…”

“…Fortunately, the CRISPR/Cas system makes it possible to establish a direct linear positive correlation between mycotoxins and fluorescence signals. − The CRISPR-Cas system can achieve binding to a matched RNA or DNA by designing specific guide RNA strands, becoming an ideal molecular detection tool based on this feature . The CRISPR/Cas12a system is based on the Cas12a enzyme to indiscriminately cleave surrounding single-stranded DNA (ssDNA) probes when the functional DNA encounters the target analyte. , Furthermore, Cas12a has low intrinsic tolerance for crRNA-target DNA mismatches and requires high complementarity, which makes the Cas12a-based detection method have high specificity . In addition, the sensing is not constrained on the interface space, thereby significantly improving the recognition efficiency.…”