2019
DOI: 10.1002/ange.201910772
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Exploring the Trans‐Cleavage Activity of CRISPR‐Cas12a (cpf1) for the Development of a Universal Electrochemical Biosensor

Abstract: An accurate, rapid, and cost‐effective biosensor for the quantification of disease biomarkers is vital for the development of early‐diagnostic point‐of‐care systems. The recent discovery of the trans‐cleavage property of CRISPR type V effectors makes CRISPR a potential high‐accuracy bio‐recognition tool. Herein, a CRISPR‐Cas12a (cpf1) based electrochemical biosensor (E‐CRISPR) is reported, which is more cost‐effective and portable than optical‐transduction‐based biosensors. Through optimizing the in vitro tran… Show more

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Cited by 123 publications
(124 citation statements)
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“…Several sensor types have been created by researchers so far, in which no target-amplification is needed to measure at a very low LOD. These sensors are a graphene-based field effect transistor (gFET) ( Hajian et al, 2019 ), nanopore sensors ( Weckman et al, 2019 ; Yang et al, 2018 ), electrochemical sensors ( Bruch et al, 2019 ; Dai et al, 2019 ; Xu et al, 2020 ; D. Zhang et al, 2020 ) and a conductivity sensor combined with a DNA gel ( English et al, 2019 ).…”
Section: Crispr/cas Sensing In Poc Sensorsmentioning
confidence: 99%
See 1 more Smart Citation
“…Several sensor types have been created by researchers so far, in which no target-amplification is needed to measure at a very low LOD. These sensors are a graphene-based field effect transistor (gFET) ( Hajian et al, 2019 ), nanopore sensors ( Weckman et al, 2019 ; Yang et al, 2018 ), electrochemical sensors ( Bruch et al, 2019 ; Dai et al, 2019 ; Xu et al, 2020 ; D. Zhang et al, 2020 ) and a conductivity sensor combined with a DNA gel ( English et al, 2019 ).…”
Section: Crispr/cas Sensing In Poc Sensorsmentioning
confidence: 99%
“…7 An overview of the mechanisms in the different electronic sensors described in the literature. A) gFET ( Hajian et al, 2019 ), B) nanopore sensor, adjusted from ( Yang et al, 2018 ), C) different types of electrochemical sensors with MB as an electrochemical tag, based on targeted (II) and collateral cleavage (I & III) ( Dai et al, 2019 ; Xu et al, 2020 ; D. Zhang et al, 2020 ) D) collateral cleavage based sensor with a glucose oxidase electrochemical tag. Adapted from ( Bruch et al, 2019 ), according to Creative Commons Attribution License (CC BY 4.0) E) collateral cleavage of DNA-gel based sensor with combined colorimetric and amperometric readout ( English et al, 2019 ).…”
Section: Crispr/cas Sensing In Poc Sensorsmentioning
confidence: 99%
“…Overall, LbCas12a showed the highest fluorescence signal, which is consistent with previous studies. 13,18 Through observation of the fluorophore-quencher-based reporter assay and time-dependent gel electrophoresis, we hypothesized that the various extensions of ssDNA on the crRNA induce conformational changes on LbCas12a that result in enhanced endonuclease activity.…”
mentioning
confidence: 99%
“…One of the many examples of successful identification of CRISPR proteins with previously unknown properties is Cas13a (previously referred to as C2c2), which has transformed into a powerful molecular tool 16 . Application of CRISPR may go far beyond detection of nucleic acids, as several CRISPR technologies have been devised to identify proteins 66 and could be utilized for detecting small molecules 111 (e.g. toxic agents, opioids, drugs etc.).…”
Section: Resultsmentioning
confidence: 99%
“…Dai and colleagues created E-CRISPR system is equipped with electrochemical signal detection module utilizing Cas12a trans-cleavage activity and modified ssDNA reporter with methylene blue electrochemical tag attached to the sensor surface 66 . In the presence of target sequence, Cas12a proteins cleave ssDNA-MB reporter decreasing the level of electrochemical signal being transduced.…”
Section: Methodsmentioning
confidence: 99%