A CRISPR-derived biosensor for the sensitive detection of transcription factors based on the target-induced inhibition of Cas12a activation

Li, Bingzhi; Xia, Anqi; Zhang, Shilin; Suo, Tiying; Ma, Yujie; Huang, He; Xing, Zhou; Chen, Yue; Zhou, Xuemin

doi:10.1016/j.bios.2020.112619

Cited by 35 publications

(11 citation statements)

References 46 publications

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“…It has been already demonstrated to not only digest target double-strand DNA (dsDNA) (named cis-cleavage) but also digest any nontarget-ssDNA without discrimination (named trans-cleavage), which leads to the superpotent lateral branch cleavage efficiency (at least 10 3 turnovers per second). In a typical splitting decomposition procedure, it can swiftly recognize the protospacer adjacent motif (PAM) , rich in T nucleotides and catalyze the maturation of the CRISPR RNA (crRNA) , to connect with activator DNA (acDNA), stimulating the trans-cleavage activity of the effector CRISPR/Cas12a protein to digest adjacent nontarget-ssDNA reporters indiscriminately. When dsDNA is present in biological components as a substrate with a reporter system, the side branch cleavage effect of CRISPR/Cas12a will directly transduct the ECL signal output, so as to achieve the goal of signal conversion.…”

Section: Introductionmentioning

confidence: 99%

3D DNA Walker-Assisted CRISPR/Cas12a Trans-Cleavage for Ultrasensitive Electrochemiluminescence Detection of miRNA-141

et al. 2021

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In this study, a CRISPR/Cas12a (LbCpf1)mediated electrochemiluminescence (ECL) paper-based platform on the basis of a three-dimensional (3D) DNA walker was proposed for the ultrasensitive detection of miRNA-141. Initially, 3D-rGO with a tremendous loading space was modified on the paper working electrode (PWE) to construct an excellent conductive substrate and facilitate the growth of AuPd nanoparticles (NPs). Afterward, the AuPd NPs were introduced as the coreaction emitter medium of the 3D-rGO/PWE to provide convenience for the transformation between S 2 O 8 2− and SO 4 2− , amplifying the ECL emission of g-C 3 N 4 nanosheets (NSs). Meanwhile, with the help of Nt.BsmAI nicking endonuclease, a 3D DNA walker signal amplifier was designed to convert and magnify the target miRNA-141 into a particular trigger sequence, which could act as activator DNA to motivate the trans-acting deoxyribonuclease activity of CRISPR/Cas12a to further achieve efficient annihilation of the ECL signal. Furthermore, the proposed multimechanism-driven biosensor exhibited excellent sensitivity and specificity, with a relatively low detection limit at 0.331 fM (S/ N = 3) in the concentration range between 1 fM and 10 nM. Consequently, the designed strategy not only extended the application scope of CRISPR/Cas12a but also devoted a new approach for the clinical diagnosis of modern medicine.

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Section: Introductionmentioning

confidence: 99%

3D DNA Walker-Assisted CRISPR/Cas12a Trans-Cleavage for Ultrasensitive Electrochemiluminescence Detection of miRNA-141

et al. 2021

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“…21,22 Afterward, newer methods were introduced by utilizing the sequence-nonspecific collateral cleavage activity found in Cas12 and Cas13 proteins that has facilitated the development of numerous diagnostic tools for rapid and portable detection, consequently yielding great potential for point of care settings. 23–26 Beyond nucleic acid detection, 27–29 Cas12a is being recently exploited to construct novel strategies for the detection of various biological analytes such as small molecules, 30 lipopolysaccharide, 31 transcription factors, 32 and enzyme activities 33 due to its exceptional capability for signal amplification by cleaving nearby ssDNA reporter upon target DNA recognition.…”

mentioning

confidence: 99%

CRISPR/Cas12a collateral cleavage activity for an ultrasensitive assay of RNase H

Kim

Lee

et al. 2022

Chem. Commun.

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“…Recently, researchers have developed a variety of approaches to detect ions [ 27 ], small molecules [ 27 , 28 ], nucleic acids [ [29] , [30] , [31] , [32] , [33] ], and transcriptional factors [ 34 ] (proteins [ 33 ]) based on the assistance of CRISPR-Cas12a. Xiong et al [ 27 ] reported a versatile detection tool combining functional DNA and Cas12a, which harbored the “competition” idea as well.…”

Section: Discussionmentioning

confidence: 99%

Aptamer-based cell-free detection system to detect target protein

Chen

Zhuang

Zheng

et al. 2021

Synthetic and Systems Biotechnology

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Biomarkers of disease, especially protein, show great potential for diagnosis and prognosis. For detecting a certain protein, a binding assay implementing antibodies is commonly performed. However, antibodies are not thermally stable and may cause false-positive when the sample composition is complicated. In recent years, a functional nucleic acid named aptamer has been used in many biochemical analysis cases, which is commonly selected from random sequence libraries by using the systematic evolution of ligands by exponential enrichment (SELEX) techniques. Compared to antibodies, the aptamer is more thermal stable, easier to be modified, conjugated, and amplified. Herein, an A ptamer- B ased C ell-free D etection (ABCD) system was proposed to detect target protein, using epithelial cell adhesion molecule (EpCAM) as an example. We combined the robustness of aptamer in binding specificity with the signal amplification ability of CRISPR-Cas12a′s trans -cleavage activity in the ABCD system. We also demonstrated that the ABCD system could work well to detect target protein in a relatively low limit of detection (50–100 nM), which lay a foundation for the development of portable detection devices. This work highlights the superiority of the ABCD system in detecting target protein with low abundance and offers new enlightenment for future design and development.

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A CRISPR-derived biosensor for the sensitive detection of transcription factors based on the target-induced inhibition of Cas12a activation

Cited by 35 publications

References 46 publications

3D DNA Walker-Assisted CRISPR/Cas12a Trans-Cleavage for Ultrasensitive Electrochemiluminescence Detection of miRNA-141

3D DNA Walker-Assisted CRISPR/Cas12a Trans-Cleavage for Ultrasensitive Electrochemiluminescence Detection of miRNA-141

CRISPR/Cas12a collateral cleavage activity for an ultrasensitive assay of RNase H

Aptamer-based cell-free detection system to detect target protein

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