CRISPR/Cas12a-based biosensor for colorimetric detection of serum prostate-specific antigen by taking nonenzymatic and isothermal amplification

Wang, Wanhe; Liu, Jingqi; Li, Xiaoxia; Lin, Chuankai; Wang, Xueliang; Liu, Jianhua; Ling, Liansheng; Wang, Jing

doi:10.1016/j.snb.2021.131228

Cited by 48 publications

(12 citation statements)

References 49 publications

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“…For example, AuNPs coupled with Cas12a collateral digestion and RCA amplification were used for colorimetric CRISPR-Cas sensing, where aptamer/crRNA/Cas12a ternary complexes cleave primer sequences and padlock probes modified on AuNPs. Wang et al [ 69 ] used distance-dependent optical properties of AuNPs and nicking enzyme-free amplification to produce more acDNA and detected aflatoxin M1 (AFM1) with ppb level of accuracy and sensing of serum PSA [ 69 , 70 ].…”

Section: Colorimetry-based Sensingmentioning

confidence: 99%

Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets

et al. 2023

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CRISPR-Cas systems have been widely used in genome editing and transcriptional regulation. Recently, CRISPR-Cas effectors are adopted for biosensor construction due to its adjustable properties, such as simplicity of design, easy operation, collateral cleavage activity, and high biocompatibility. Aptamers’ excellent sensitivity, specificity, in vitro synthesis, base-pairing, labeling, modification, and programmability has made them an attractive molecular recognition element for inclusion in CRISPR-Cas systems. Here, we review current advances in aptamer-based CRISPR-Cas sensors. We briefly discuss aptamers and the knowledge of Cas effector proteins, crRNA, reporter probes, analytes, and applications of target-specific aptamers. Next, we provide fabrication strategies, molecular binding, and detection using fluorescence, electrochemical, colorimetric, nanomaterials, Rayleigh, and Raman scattering. The application of CRISPR-Cas systems in aptamer-based sensing of a wide range of biomarkers (disease and pathogens) and toxic contaminants is growing. This review provides an update and offers novel insights into developing CRISPR-Cas-based sensors using ssDNA aptamers with high efficiency and specificity for point-of-care setting diagnostics.

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Section: Colorimetry-based Sensingmentioning

confidence: 99%

Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets

et al. 2023

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“…CRISPR/Cas12a colorimetric biosensor 0.2-40 ng/mL 0.1 ng/mL [37] Aptamer-based frequency shift Raman sensing 0.05-25 ng/mL 10 pg/mL [38] Ratiometric antifouling electrochemical biosensor 0.005-10 ng/mL 0.83 pg/mL [39] Magnetic-assisted Bi 2 MoO 6 -based PEC biosensor 0.005-100 ng/mL 3.5 pg/mL [40] BiOClÀ Au/FTO-based PEC immunoassay 0.01-50 ng/mL 2.3 pg/mL This work the aforementioned concentrations, respectively, demonstrating good reproducibility. The specificity of the split-type BiOClÀ Au/FTO-based PEC immunoassay was evaluated by assaying other cancer biomarkers as interference agents (50 ng/mL), e. g., human epidermal growth factor receptor 2 (HER2), carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) under the same experimental conditions.…”

Section: Signal Transduction Strategymentioning

confidence: 99%

Surface plasmon resonance‐enhanced photoelectrochemical immunoassay of prostate‐specific antigen based on BiOCl‐Au heterojunction

et al. 2023

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Taking advantage of the high-efficiency photocurrent response of bismuth oxychloride sensitized with gold nanoparticles (BiOClÀ Au) in combination with the antigen-antibody biological recognition reaction, a convenient photoelectrochemical (PEC) immunoassay (model target: prostate-specific antigen; PSA) has been fabricated. In particular, the enhanced photocurrent response could be attributed to the enhanced optical absorption of visible light from surface plasmon resonance (SPR) effect of gold nanoparticles in the hierarchical structure of BiOCl layered. To realize the biological detection process, an immunoreaction was implemented between target PSA and alkaline phosphatase (ALP)-labeled anti-PSA antibodies. With the formation of ternary sandwich immunocomplex, the carried and loaded ALP catalysed the substrate ascorbic acid 2-phosphate (AAP) to generate ascorbic acid for increasing the photocurrent intensity of BiOClÀ Au/FTO. Under optimum reaction time, the photocurrent peak intensity of BiOClÀ Au/FTO increased with the increasing of target PSA concentration in the range of 0.01 ng/mL to 50 ng/mL with a limit of detection (LOD) down to 2.3 pg/mL. In the photocurrent control experiment, the photocurrent of BiOClÀ Au/FTO was not only higher than that of BiOCl/FTO, but also showed greater changes in photocurrent under the same target PSA concentration. Impressively, the proposed split-type PEC immunoassay was applied to detect PSA concentration in human serum samples, giving acceptable and satisfactory accuracy compared with the gold standard PSA ELISA method.

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“…In this work, the exonuclease polymerase is harnessed for releasing cDNA from HP-cDNA during SDA, along with triggering the next SDA cycle, this unique design renders the biosensor simpler and more convenient for clinical testing. The same group also reported a colorimetric assay for serum PSA using the nonenzymatic and isothermal properties of HCR to convert serum PSA into nucleic acid products [ 83 ]. The presence of PSA triggers HCR amplification to produce dsDNA containing multiple PAM sites recognized by Cas12a, activating Cas12a’s trans cleavage activity, which nonspecifically cleaves the DNA–AuNP probe pairs along with a colorimetric signal.…”

Section: Aptamer-assisted Crispr/cas-based Protein Detectionmentioning

confidence: 99%

Section: Aptamer-assisted Crispr/cas-based Protein Detectionmentioning

confidence: 99%

Recent Advances in CRISPR/Cas-Based Biosensors for Protein Detection

et al. 2022

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CRISPR is an acquired immune system found in prokaryotes that can accurately recognize and cleave foreign nucleic acids, and has been widely explored for gene editing and biosensing. In the past, CRISPR/Cas-based biosensors were mainly applied to detect nucleic acids in the field of biosensing, and their applications for the detection of other types of analytes were usually overlooked such as small molecules and disease-related proteins. The recent work shows that CRISPR/Cas biosensors not only provide a new tool for protein analysis, but also improve the sensitivity and specificity of protein detections. However, it lacks the latest review to summarize CRISPR/Cas-based biosensors for protein detection and elucidate their mechanisms of action, hindering the development of superior biosensors for proteins. In this review, we summarized CRISPR/Cas-based biosensors for protein detection based on their mechanism of action in three aspects: antibody-assisted CRISPR/Cas-based protein detection, aptamer-assisted CRISPR/Cas-based protein detection, and miscellaneous CRISPR/Cas-based methods for protein detection, respectively. Moreover, the prospects and challenges for CRISPR/Cas-based biosensors for protein detection are also discussed.

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CRISPR/Cas12a-based biosensor for colorimetric detection of serum prostate-specific antigen by taking nonenzymatic and isothermal amplification

Cited by 48 publications

References 49 publications

Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets

Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets

Surface plasmon resonance‐enhanced photoelectrochemical immunoassay of prostate‐specific antigen based on BiOCl‐Au heterojunction

Recent Advances in CRISPR/Cas-Based Biosensors for Protein Detection

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