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

Wang, Jing; Yang, Xiaorong; Wang, Xueliang; Wang, Wanhe

doi:10.3390/bioengineering9100512

Cited by 12 publications

(7 citation statements)

References 126 publications

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“…State-of-the-art electrochemical bioplatforms have been developed by using CRISPR/Cas systems as biorecognition elements for single or multiplexed determination of nucleic acids, or as signal amplifiers to interrogate nucleic acids or other analytes such as proteins, metals, and bacteria. − Indeed, the versatility of modern electrochemical platforms and instrumentation involving CRISPR/Cas technologies allow the determination of markers of different molecular levels and the development of multiplexed electrochemical bioplatforms.…”

Section: Key Alliances To Cover Important Routesmentioning

confidence: 99%

Electrochemical Affinity Biosensors: Pervasive Devices with Exciting Alliances and Horizons Ahead

Campuzano

Pingarrón

2023

ACS Sens.

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Electrochemical affinity biosensors are evolving at breakneck speed, strengthening and colonizing more and more niches and drawing unimaginable roadmaps that increasingly make them protagonists of our daily lives. They achieve this by combining their intrinsic attributes with those acquired by leveraging the significant advances that occurred in (nano)materials technology, bio(nano)materials and nature-inspired receptors, gene editing and amplification technologies, and signal detection and processing techniques. The aim of this Perspective is to provide, with the support of recent representative and illustrative literature, an updated and critical view of the repertoire of opportunities, innovations, and applications offered by electrochemical affinity biosensors fueled by the key alliances indicated. In addition, the imminent challenges that these biodevices must face and the new directions in which they are envisioned as key players are discussed.

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Section: Key Alliances To Cover Important Routesmentioning

confidence: 99%

Electrochemical Affinity Biosensors: Pervasive Devices with Exciting Alliances and Horizons Ahead

Campuzano

Pingarrón

2023

ACS Sens.

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“…LbCas12a protein binding to Protospacer adjacent motif directly leads to local melting of PAM downstream target sequence DNA, causing downstream sequence complementation with speci cally recognized gRNA, thereby activating cleavage of target DNA by Cas12a protein [23]. The activity of the Cas12a protein directly affects the detection e cacy of the CRISPR/Cas12a system.…”

Section: Validation Of Cleavage Activity Of Lbcas12a Proteinmentioning

confidence: 99%

A new method for the detection of Mycobacterium tuberculosis based on the CRISPR/Cas system

Zhang

et al. 2023

Preprint

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Object: Mycobacterium tuberculosis (MTB) is a bacterium that can cause zoonoses by aerosol transmission. Tuberculosis (TB) caused by MTB places a heavy burden on world public health security. The development of efficient, specific, convenient, and inexpensive MTB assays is important for the prevention and control of TB. Methods: Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) is a special DNA repeat family widely present in bacterial and archaeal genomes. In this study, we established a specific detection method for MTB using the CRISPR system, combined with recombinase mediated isothermal nucleic acid amplification (RAA) to improve the sensitivity of the detection system and achieve "two-level" amplification of the detection signal. The sensitivity and specificity of RAA combined with the CRISPR/Cas system were analyzed. Using BACTEC 960 culture as the gold standard for the detection of MTB, we established the TB-CRISPR technique by testing 504 samples from patients with suspected tuberculosis. Results: MTB H37Ra could be detected as low as 3.13 CFU/mL by the CRISPR-Cas12a system targeting IS6110. With BACTCE960 culture (120 positives and 384 negatives) as the gold standard, the sensitivity of the TB-CRISPR technique was 0.883 (0.809-0.932) and the specificity was 0.940 (0.910-0.961). According to the receiver operating characteristic (ROC) curve analysis, the area under the curve (AUC) reached 0.944 (0.914-0.975) within 95% CI. The positive likelihood ratio (PLR) was 14.747 (9.870-22.035) and the negative likelihood ratio (NLR) was 0.124 (0.076-0.203). The positive predictive value (PPV) was 0.822 (0.742-0.881) and the negative predictive value (NPV) was 0.963 (0.937-0.979). Conclusion: TB-CRISPR realizes the rapid screening and diagnosis of MTB. The whole detection time is less than 1.5h. It is easy to operate and does not need to rely on complex instruments. It is of great significance for the rapid detection of MTB and the clinical diagnosis of TB.

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“…The results showed that the fluorescence bioassay based on CRISPR-Cas12a is able to accurately distinguish people with cancer from healthy ones. [189]…”

Section: Crispr As a Tool For Diagnosismentioning

confidence: 99%

“…They found that the CRISPR‐Cas12a‐based fluorescence bioassay against mismatched sequences had a high specificity as well as a high sensitivity of about 161 fM for the determination of miR‐21. The results showed that the fluorescence bioassay based on CRISPR‐Cas12a is able to accurately distinguish people with cancer from healthy ones [189] …”

Section: Crispr‐cas 9 Systems: Components and Limitationsmentioning

confidence: 99%

Recent Progress in Prompt Molecular Detection of Exosomes Using CRISPR/Cas and Microfluidic‐Assisted Approaches Toward Smart Cancer Diagnosis and Analysis

Mojtaba Mousavi,

Alireza Hashemi,

Yari Kalashgrani

et al. 2023

ChemMedChem

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Exosomes are essential indicators of molecular mechanisms involved in interacting with cancer cells and the tumor environment. As nanostructures based on lipids and nucleic acids, exosomes provide a communication pathway for information transfer by transporting biomolecules from the target cell to other cells. Importantly, these extracellular vesicles are released into the bloodstream by the most invasive cells, i. e., cancer cells; in this way, they could be considered a promising specific biomarker for cancer diagnosis. In this matter, CRISPR‐Cas systems and microfluidic approaches could be considered practical tools for cancer diagnosis and understanding cancer biology. CRISPR‐Cas systems, as a genome editing approach, provide a way to inactivate or even remove a target gene from the cell without affecting intracellular mechanisms. These practical systems provide vital information about the factors involved in cancer development that could lead to more effective cancer treatment. Meanwhile, microfluidic approaches can also significantly benefit cancer research due to their proper sensitivity, high throughput, low material consumption, low cost, and advanced spatial and temporal control. Thereby, employing CRISPR‐Cas‐ and microfluidics‐based approaches toward exosome monitoring could be considered a valuable source of information for cancer therapy and diagnosis. This review assesses the recent progress in these promising diagnosis approaches toward accurate cancer therapy and in‐depth study of cancer cell behavior.

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

Cited by 12 publications

References 126 publications

Electrochemical Affinity Biosensors: Pervasive Devices with Exciting Alliances and Horizons Ahead

Electrochemical Affinity Biosensors: Pervasive Devices with Exciting Alliances and Horizons Ahead

A new method for the detection of Mycobacterium tuberculosis based on the CRISPR/Cas system

Recent Progress in Prompt Molecular Detection of Exosomes Using CRISPR/Cas and Microfluidic‐Assisted Approaches Toward Smart Cancer Diagnosis and Analysis

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