CRISPR-Cas12a Coupled with Platinum Nanoreporter for Visual Quantification of SNVs on a Volumetric Bar-Chart Chip

Shao, Ning; Han, Xin; Song, Yanni; Zhang, Pengchao; Qin, Lidong

doi:10.1021/acs.analchem.9b02925

Cited by 119 publications

(96 citation statements)

References 30 publications

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“…Shao et al proposed a visual and stand-alone SNP detection method based on CRISPR/Cas12a cleavage ( Shao et al, 2019 ). By their volumetric bar-chart chip, multiple cancer mutations could be identified and quantified in serum.…”

Section: Crispr/cas Sensing In Poc Sensorsmentioning

confidence: 99%

“…So far only two multiplexed biosensing systems were published based on CRISPR/Cas systems ( Gootenberg et al, 2018 ; Shao et al, 2019 ). Li et al describe three different strategies for in vitro based multiplexed CRISPR sensing: Cas effector-based, signal transduction-based and separation-based ( Y. Li et al, 2019b ).…”

Section: Challenges In Poc Crispr Sensing Systemsmentioning

confidence: 99%

“…By physical separation Shao et al achieved multiplex CRISPR sensing by separating Cas effector proteins in different microfluidic channels. In this way each channel is used to test a different sequence by changing the spacer sequence on the crRNA ( Shao et al, 2019 ). This microfluidic approach, using which spatial separation can be achieved, is not limited by the current status of the CRISPR biotechnology or advanced transducers for read-out.…”

Section: Challenges In Poc Crispr Sensing Systemsmentioning

confidence: 99%

See 2 more Smart Citations

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

Dongen

Berendsen

Steenbergen

et al. 2020

Biosensors and Bioelectronics

284

191

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With the trend of moving molecular tests from clinical laboratories to on-site testing, there is a need for nucleic acid based diagnostic tools combining the sensitivity, specificity and flexibility of established diagnostics with the ease, cost effectiveness and speed of isothermal amplification and detection methods. A promising new nucleic acid detection method is Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas)-based sensing. In this method Cas effector proteins are used as highly specific sequence recognition elements that can be combined with many different read-out methods for on-site point-of-care testing. This review covers the technical aspects of integrating CRISPR/Cas technology in miniaturized sensors for analysis on-site. We start with a short introduction to CRISPR/Cas systems and the different effector proteins and continue with reviewing the recent developments of integrating CRISPR sensing in miniaturized sensors for point-of-care applications. Finally, we discuss the challenges of point-of-care CRISPR sensing and describe future research perspectives.

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Section: Crispr/cas Sensing In Poc Sensorsmentioning

confidence: 99%

Section: Challenges In Poc Crispr Sensing Systemsmentioning

confidence: 99%

Section: Challenges In Poc Crispr Sensing Systemsmentioning

confidence: 99%

See 1 more Smart Citation

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

Dongen

Berendsen

Steenbergen

et al. 2020

Biosensors and Bioelectronics

284

191

View full text Add to dashboard Cite

show abstract

“…Designs realizing spatial or local separation offer another etection strategy, since CRISPR‐Cas12a systems can effectively operated both in solution and on solid‐based surfaces. Devices integrating separated Cas12a/crRNA binary complexes targeting various targets have enabled separation‐based approaches, which can operate multiple separated micro‐volume reactions restricted in small zones, such as hydrogels, [12] paper‐based microfluidic devices, [12] lateral flow test strips [10b,f,32d,e,33a,41] or volumetric bar‐chart chips [25g] . The methods integrating the Cas12a system and separation design have enabled highly multiplexed detection at lower cost by reducing the reaction volume.…”

Section: The Application Of Crispr‐cas12a In Biosensingmentioning

confidence: 99%

CRISPR‐Cas12a System for Biosensing and Gene Regulation

Shi

Yao

et al. 2021

Chemistry An Asian Journal

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Clustered regularly interspaced short palindromic repeats (CRISPR) is a promising technology in the biological world. As one of the CRISPR-associated (Cas) proteins, Cas12a is an RNA-guided nuclease in the type V CRISPR-Cas system, which has been a robust tool for gene editing. In addition, due to the discovery of target-binding-induced indiscriminate single-stranded DNase activity of Cas12a, CRISPR-Cas12a also exhibits great promise in biosensing. This minireview not only gives a brief introduction to the mechanism of CRISPR-Cas12a but also highlights the recent developments and applications in biosensing and gene regulation. Finally, future prospects of the CRISPR-Cas12a system are also discussed. We expect this minireview will inspire innovative work on the CRISPR-Cas12a system by making full use of its features and advantages.

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“…For instance, with the mediation of CRISPR RNA (crRNA), Cas12a can recognize specific DNA sequences and exhibit nonspecific single‐stranded DNA (ssDNA) cleavage activity (Swarts & Jinek, 2019; Yamano et al, 2016). Based on such trans‐cleavage activities, CRISPR Cas12a has been transformed into several biosensors for the detection of nucleic acid targets, such as single‐nucleotide polymorphism (Li et al, 2018; Shao et al, 2019), pathogenic bacteria (Chen et al, 2020; Wang et al, 2020), bacterial resistances (English et al, 2019; Xu et al, 2020), and viruses (Chen et al, 2018; Tsou et al, 2019). What's more, several biosensors for sensitive detection of SARS‐CoV‐2 have been established by combining CRISPR Cas12a with isothermal amplification methods, such as reverse transcription loop‐mediated isothermal amplification and recombinase polymerase amplification (RPA).…”

Section: Introductionmentioning

confidence: 99%

A CRISPR‐Cas12a‐derived biosensor enabling portable personal glucose meter readout for quantitative detection of SARS‐CoV‐2

Huang

Shi

Qian

et al. 2021

Biotech & Bioengineering

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Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has spread rapidly throughout the whole world and caused significant difficulties in the prevention and control of the epidemic. In this case, several detection methods have been established based on nucleic acid diagnostic techniques and immunoassays to achieve sensitive and specific detection of SARS‐CoV‐2. However, most methods are still largely dependent on professional instruments, highly trained operators, and centralized laboratories. These limitations gravely diminish their practicality and portability. Herein, a clustered regularly interspaced short palindromic repeats (CRISPR) Cas12a based assay was developed for portable, rapid and sensitive of SARS‐CoV‐2. In this assay, samples were quickly pretreated and amplified by reverse transcription recombinase‐aided amplification under mild conditions. Then, by combining the CRISPR Cas12a system and a glucose‐producing reaction, the signal of the virus was converted to that of glucose, which can be quantitatively read by a personal glucose meter in a few seconds. Nucleocapsid protein gene was tested as a model target, and the sensitivity for quantitative detection was as low as 10 copies/μl, which basically meet the needs of clinical diagnosis. In addition, with the advantages of lower material cost, shorter detection time, and no requirement for professional instrument in comparison with quantitative reverse transcription‐polymerase chain reaction, this assay is expected to provide a powerful technical support for the early diagnosis and intervention during epidemic prevention and control.

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CRISPR-Cas12a Coupled with Platinum Nanoreporter for Visual Quantification of SNVs on a Volumetric Bar-Chart Chip

Cited by 119 publications

References 30 publications

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

CRISPR‐Cas12a System for Biosensing and Gene Regulation

A CRISPR‐Cas12a‐derived biosensor enabling portable personal glucose meter readout for quantitative detection of SARS‐CoV‐2

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