CRISPR/Cas12a collateral cleavage activity for simple and rapid detection of protein/small molecule interaction

Kim, Hansol; Lee, Seo-Young; Yoon, Junhyeok; Song, Jayeon; Park, Hyun Gyu

doi:10.1016/j.bios.2021.113587

Cited by 35 publications

(25 citation statements)

References 43 publications

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“…Previous work reported that prolonging the incubation time of the reporter solution could cleave more reporters and increased the signal. 34,40 Accordingly, we measured the florescence signal and found the florescence intensity increased with the cleavage time (Figure S6). Thus, LFAs were performed again by prolonging the cleavage time from 20 to 40 min (Figure S7) and 60 min (Figure 3D).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

CRISPR-Cas12a-Based Aptasensor for On-Site and Highly Sensitive Detection of Microcystin-LR in Freshwater

Kang

et al. 2022

Environ. Sci. Technol.

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On-site monitoring of trace organic pollutants with facile methods is critical to environmental pollutant prevention and control. Herein, we proposed a CRISPR-Cas12a-based aptasensor platform (named as MC-LR-Casor) for on-site and sensitive detection of microcystin-LR (MC-LR). After hybridization with blocker DNA, the MC-LR aptamers were conjugated to magnetic beads (MBs) to get the MB aptasensor. In the presence of MC-LR, their interactions with aptamers were triggered and the specific binding caused the release of blocker DNA. Using the programmability of the CRISPR-Cas system, the released blocker DNA was designed to activate a Cas12a-crRNA complex. Single strand DNA reporters were rapidly cleaved by the complex. Signal readout could be achieved by fluorometer or lateral flow strips, which were positively correlated to MC-LR concentration. Benefiting from the CRISPR-Cas12a amplification system, the proposed sensing platform exhibited high sensitivity and reached the limit of detection of ∼3 × 10–6 μg/L (fluorescence method) or 1 × 10–3 μg/L (lateral flow assay). In addition, the MC-LR-Casor showed excellent selectivity and good recovery rates, demonstrating their good applicability for real water sample analysis. During the whole assay, only two steps of incubation at a constant temperature were required and the results could be visualized when employing flow strips. Therefore, the proposed assay offered a simple and convenient alternative for in situ MC-LR monitoring, which may hold great promise for future environmental surveillance.

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Section: ■ Results and Discussionmentioning

confidence: 99%

CRISPR-Cas12a-Based Aptasensor for On-Site and Highly Sensitive Detection of Microcystin-LR in Freshwater

Kang

et al. 2022

Environ. Sci. Technol.

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“…Additionally, with the adoption of AuNP, the applications of CRISPR-driven biosensors were potentially extended, such as the ultrasensitive detection of mycotoxins including aflatoxin M1 ( Abnous et al, 2021 ), assessment for telomerase activity by analyzing telomeric repeat DNA, and internal control products ( Cheng et al, 2021 ). Furthermore, CRISPR/Cas12a-based biosensors can monitor protein/small molecule interactions, like streptavidin/biotin and anti-digoxigenin/digoxigenin ( Kim et al, 2021 ).…”

Section: Applications Of Crispr-based Biosensing Techniquesmentioning

confidence: 99%

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

Chen

et al. 2022

Front. Bioeng. Biotechnol.

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In the fight against the worldwide pandemic coronavirus disease 2019 (COVID-19), simple, rapid, and sensitive tools for nucleic acid detection are in urgent need. PCR has been a classic method for nucleic acid detection with high sensitivity and specificity. However, this method still has essential limitations due to the dependence on thermal cycling, which requires costly equipment, professional technicians, and long turnover times. Currently, clustered regularly interspaced short palindromic repeats (CRISPR)-based biosensors have been developed as powerful tools for nucleic acid detection. Moreover, the CRISPR method can be performed at physiological temperature, meaning that it is easy to assemble into point-of-care devices. Microfluidic chips hold promises to integrate sample processing and analysis on a chip, reducing the consumption of sample and reagent and increasing the detection throughput. This review provides an overview of recent advances in the development of CRISPR-based biosensing techniques and their perfect combination with microfluidic platforms. New opportunities and challenges for the improvement of specificity and efficiency signal amplification are outlined. Furthermore, their various applications in healthcare, animal husbandry, agriculture, and forestry are discussed.

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“…A small molecule modification strategy for fDNA has been reported in recent studies (Kim et al, 2021). Small molecules can serve as key components for the recognition of target proteins, and the presence or absence of complexes composed of target proteins and small molecules affects the binding efficiency of crRNA during downstream signaling.…”

Section: Non-nucleic-acid Detection Strategies 241 Based On Fdna Apta...mentioning

confidence: 99%

Research progress of CRISPR-based biosensors and bioassays for molecular diagnosis

Chen

Shen

Wang

et al. 2022

Front. Bioeng. Biotechnol.

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CRISPR/Cas technology originated from the immune mechanism of archaea and bacteria and was awarded the Nobel Prize in Chemistry in 2020 for its success in gene editing. Molecular diagnostics is highly valued globally for its development as a new generation of diagnostic technology. An increasing number of studies have shown that CRISPR/Cas technology can be integrated with biosensors and bioassays for molecular diagnostics. CRISPR-based detection has attracted much attention as highly specific and sensitive sensors with easily programmable and device-independent capabilities. The nucleic acid-based detection approach is one of the most sensitive and specific diagnostic methods. With further research, it holds promise for detecting other biomarkers such as small molecules and proteins. Therefore, it is worthwhile to explore the prospects of CRISPR technology in biosensing and summarize its application strategies in molecular diagnostics. This review provides a synopsis of CRISPR biosensing strategies and recent advances from nucleic acids to other non-nucleic small molecules or analytes such as proteins and presents the challenges and perspectives of CRISPR biosensors and bioassays.

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CRISPR/Cas12a collateral cleavage activity for simple and rapid detection of protein/small molecule interaction

Cited by 35 publications

References 43 publications

CRISPR-Cas12a-Based Aptasensor for On-Site and Highly Sensitive Detection of Microcystin-LR in Freshwater

CRISPR-Cas12a-Based Aptasensor for On-Site and Highly Sensitive Detection of Microcystin-LR in Freshwater

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

Research progress of CRISPR-based biosensors and bioassays for molecular diagnosis

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