CRISPR-Cas-mediated diagnostics

Li, Luoxi; Shen, Guanghui; Wu, Min; Jiang, Jianxin; Xia, Qingyou; Lin, Ping

doi:10.1016/j.tibtech.2022.04.006

Cited by 43 publications

(13 citation statements)

References 120 publications

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“…Due to its ability to accurately recognize and cut specific DNA and RNA sequences, the CRISPR/Cas system has been used for genome editing. , Additionally, upon recognition of a target sequence, certain CRISPR/Cas systems (including homologues of Cas13, Cas12a, and Cas14) exhibit concomitant nonspecific catalytic activity. , This cleavage activity can be used for nucleic acid detection: for example, Cas12a can degrade labeled reporter probes to generate fluorescent signals. , The CRISPR/Cas system is capable of recognizing target molecule concentrations as low as attomolar (10 –18 mol/L). Integrating CRISPR/Cas with LFSBs enables inexpensive, accurate, and highly sensitive POCT for various targets including virus, noncoding RNA, and pathogenic bacteria. − Traditional nucleic acid LFSBs usually use LAMP or RPA for nucleic acid amplification, which is prone to false positives caused by primers and primer dimers. Since a specific sequence in the target double-stranded DNA can activate the trans-cleavage activity of Cas12a in the CRISPR/Cas system, the CRISPR/Cas12a-based LFSB greatly improves the detection accuracy.…”

Section: Lfsbs For Indirect Detection Of Foodborne Pathogenic Bacteriamentioning

confidence: 99%

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review

Huang

Cheng

et al. 2023

J. Agric. Food Chem.

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Rapid and sensitive detection of foodborne pathogenic bacteria is particularly important for the prevention and control of foodborne diseases. The lateral flow strip biosensor (LFSB) is one of the most promising point-of-care detection tools and has been widely used in food safety monitoring. This review introduces recent advances in the detection of foodborne pathogenic bacteria using LFSBs. According to different bacterial biomarkers, we summarize the direct and indirect sensing strategies of bacterial LFSBs. The direct sensing strategies for whole bacterial cells are divided into antibodies, antibody alternatives, and label-free according to the recognition elements. The indirect sensing strategies refer to the detection of bacterial nucleic acids and metabolites. Next, we compare and discuss the applications of direct and indirect sensing strategies. Finally, the existing challenges, future perspectives, and development directions are discussed, which will facilitate the theoretical innovation and practical application for bacterial LFSBs.

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Section: Lfsbs For Indirect Detection Of Foodborne Pathogenic Bacteriamentioning

confidence: 99%

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review

Huang

Cheng

et al. 2023

J. Agric. Food Chem.

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“…Samples not containing the viral genetic markers will not activate Cas13; the reporter DNA remains intact and will not generate a readable signal. 13 Not only can SHERLOCK be used to detect COVID-19 or any of its variants, but it can be programmed to detect almost any virus, including the influenza virus. 14…”

Section: Diagnosticsmentioning

confidence: 99%

CRISPR Technology: A Jewish Legal Perspective

Loike¹,

Blaum²

2022

Rambam Maimonides Med J

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Clustered regularly interspaced short palindromic repeats (CRISPR) gene editing is an innovative and potentially game-changing biotechnology that can potentially reverse DNA mutations in a tissue-specific manner. In addition, CRISPR is being targeted for xenotransplantation, for increasing human longevity, in animal breeding, and in plant science. However, there are many ethical challenges that emerge from CRISPR technology. This article discusses several positions that relate to these ethical challenges from a Jewish legal perspective. In addition, we present several other applications of CRISPR technology that lack a defined Jewish legal precedent and require rabbinical scholars to address and resolve them in the future.

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“…Since the optimal temperature for Cas protease activity is similar to that of the RPA reaction, it is possible to combine the two, with the RPA responsible for thermostatically amplifying the target nucleic acid sequence and the Cas protein–crRNA complex responsible for specifically binding the target sequence and visualizing the assay results via cleaving the fluorescent reporter probe. ,, RPA-CRISPR/Cas12a assays have been used in the detection of a wide range of humans, foods, and plant pathogens including mycoplasmas, viruses, , bacteria, , and Toxoplasma gondii . However, there is no report on the application of RPA-CRISPR/Cas12a technology in the detection of fungicide resistance in plant pathogens . In the general detection platform based on RPA-CRISPR/Cas12a, a two-step method is dominant, i.e., isothermal amplification and target detection based on Cas12a are two independent steps.…”

Section: Introductionmentioning

confidence: 99%

“…30 However, there is no report on the application of RPA-CRISPR/Cas12a technology in the detection of fungicide resistance in plant pathogens. 31 In the general detection platform based on RPA-CRISPR/Cas12a, a two-step method is dominant, i.e., isothermal amplification and target detection based on Cas12a are two independent steps. Amplified products need to be transferred to the Cas12a cleavage system for target detection, which is actually inconvenient and may cause cross contamination between samples due to aerosols generated by the high amplification efficiency of RPA.…”

Section: ■ Introductionmentioning

confidence: 99%

One-Pot Assay for Rapid Detection of Benzimidazole Resistance in Venturia carpophila by Combining RPA and CRISPR/Cas12a

Liu

Cai

et al. 2023

J. Agric. Food Chem.

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High resistance to benzimidazole fungicides in Venturia carpophila is caused by the point mutation E198K of the βtubulin (TUB2) gene. Traditional methods for detection of fungicide resistance are time-consuming, which are routinely based on tedious operation, reliance on expensive equipment, and specially trained people. Therefore, it is important to establish efficient methods for field detection of benzimidazole resistance in V. carpophila to make suitable management strategies and ensure food safety. Based on recombinase polymerase amplification (RPA) combined with CRISPR/Cas12a, a rapid one-pot assay ORCas12a-BRVc (one-pot RPA-CRISPR/Cas12 platform) was established for the detection of benzimidazole resistance in V. carpophila. The ORCas12a-BRVc assay enabled one-pot detection by adding components at the bottom and wall of the tube separately, solving the problems of aerosol contamination and decreased sensitivity caused by competing DNA substrates between Cas12a cleavage and RPA amplification. The ORCas12a-BRVc assay could accomplish the detection with a minimum of 7.82 × 10 3 fg μL −1 V. carpophila genomic DNA in 45 min at 37 °C. Meanwhile, this assay showed excellent specificity due to the specific recognition ability of the Cas12a-crRNA complex. Further, we combined a method that could rapidly extract DNA from V. carpophila within 2 min with the ORCas12a-BRVc to achieve more rapid and simple detection of V. carpophila with benzimidazole resistance in fields. The ORCas12a-BRVc assay has the advantages of simplicity, rapidity, high sensitivity, high specificity, and ease of operation without the need for precision instruments and the need to isolate and culture pathogens. This assay is the first application of the one-pot platform based on the combination of RPA and CRISPR/Cas12a in fungicide resistance detection and can be used for monitoring of resistant populations in fields, providing guidance on making suitable management strategies for peach scab.

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CRISPR-Cas-mediated diagnostics

Cited by 43 publications

References 120 publications

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review

CRISPR Technology: A Jewish Legal Perspective

One-Pot Assay for Rapid Detection of Benzimidazole Resistance in Venturia carpophila by Combining RPA and CRISPR/Cas12a

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