CRISPR as a Diagnostic Tool

Kim, Seohyun; Ji, Sangmin; Koh, Hye Ran

doi:10.3390/biom11081162

Cited by 48 publications

(37 citation statements)

References 56 publications

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“…The unique region of E. fawcettii was amplified through an RPA reaction, and the reaction product was directly added to the CRISPR/Cas12a reaction. The amplicon with a unique sequence could be specifically recognized by Cas12a and subsequently triggered CRISPR/Cas12a to cleave the double-labeled ssDNA reporter through the trans-cleavage activity [ 19 , 24 ]. The lateral flow readout is based on the cleavage of a double-labeled reporter (FAM/FITC–biotin); the presence of many reporters causes anti-FAM antibody–gold nanoparticle conjugates (Au-NP) to gather at the C (control) line on the strip.…”

Section: Resultsmentioning

confidence: 99%

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Sensitive and Rapid Detection of Citrus Scab Using an RPA-CRISPR/Cas12a System Combined with a Lateral Flow Assay

Shin

Kwon

Lee

et al. 2021

Plants

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Citrus is the most extensively produced fruit tree crop in the world and is grown in over 130 countries. Fungal diseases in citrus can cause significant losses in yield and quality. An accurate diagnosis is critical for determining the best management practices and preventing future losses. In this study, a Recombinase polymerase amplification (RPA)-clustered regularly interspaced short palindromic repeats (CRISPR)/associated (Cas) system was established with the integration of a lateral flow assay (LFA) readout system for diagnosis of citrus scab. This detection can be completed within 1 h, is highly sensitive and prevents cross-reactions with other common fungal citrus diseases. Furthermore, the detection system is compatible with crude DNA extracted from infected plant tissue. This RPA-CRISPR/Cas12a-LFA system provides a sensitive, rapid, and cost-effective method with promising and significant practical value for point-of-care diagnosis of citrus scab. To our knowledge, this is the first report to establish an RPA- and CRISPR-based method with LFA for fungal diseases in plants.

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Section: Resultsmentioning

confidence: 99%

“…After cleavage, Cas12a cleaves the surrounding single-stranded DNA (ssDNA), which is also called reporter DNA, in a nonspecific manner. Combined with RPA, DETECTR has successfully been used to perform highly sensitive and specific detection of various pathogens [ 16 , 17 , 18 , 19 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

Sensitive and Rapid Detection of Citrus Scab Using an RPA-CRISPR/Cas12a System Combined with a Lateral Flow Assay

Shin

Kwon

Lee

et al. 2021

Plants

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“…The sequence-specific dsDNA binding and cleavage capabilities of the Cas9/sgRNA or dCas9/sgRNA systems have been used to develop biosensors for nucleic acid assays [ 40 , 47 , 49 ]. Because of characteristics such as adaptability, simplicity, specificity, and effectiveness, CRISPR/Cas9 technology has been extensively employed for genome editing ( Figure 3 ) and has a significant potential for biomedical studies [ 50 , 51 ].…”

Section: The Crispr/cas Systemmentioning

confidence: 99%

CRISPR/Cas: A New Tool in the Research of Telomeres and Telomerase as Well as a Novel Form of Cancer Therapy

Porika

Tippani

Saretzki

2022

IJMS

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Due to their close connection with senescence, aging, and disease, telomeres and telomerase provide a unique and vital research route for boosting longevity and health span. Despite significant advances during the last three decades, earlier studies into these two biological players were impeded by the difficulty of achieving real-time changes inside living cells. As a result of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated system’s (Cas) method, targeted genetic studies are now underway to change telomerase, the genes that govern it as well as telomeres. This review will discuss studies that have utilized CRISPR-related technologies to target and modify genes relevant to telomeres and telomerase as well as to develop targeted anti-cancer therapies. These studies greatly improve our knowledge and understanding of cellular and molecular mechanisms that underlie cancer development and aging.

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“…Метод позволяет проводить мультиплексный анализ. Чувствительность и скорость у метода довольно высокие, однако для работы Cas12a требуется PAM [25,37].…”

Section: платформы диагностики на основе систем Crispr/cas V типаunclassified

CRISPR/Cas-based diagnostic platforms

Volkov

Dolgova

Dedkov

2022

Russian Journal of Infection and Immunity

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Over the last few years, CRISPR/Cas systems have been extensively studied and used for a wide range of applied purposes. The variety of their applications is accounted for by the ability of Cas-type nucleases to targetly cleave specified nucleic acid sequences. In this case, the researcher might set the necessary sequence of the guiding elements in the CRISPR/Cas system, played by so-called single guide RNAs allowing it to act on select targets. This potential underlies one of the reasons for exerting interest in CRISPR/Cas systems. One of the first areas for applying these systems was its use for genomic editing. Later, the list of potential opportunities has been expanded: e.g., they can be used in gene therapy and epigenetic research. It is possible to create sgRNA libraries which might be used to create a pool of viral vectors applied for bacterial cell transformation with subsequent cas-protein transduction that cause target gene knockout. This approach allows finding genes responsible for resistance or sensibility to diverse substances. Using such systems in molecular diagnostics of infectious diseases is considered as one of the most promising directions allowing to detect even extremely low concentrations of pathogenic organisms in samples due to their specific nucleotide sequences. Simultaneously, such assays turn out to be accurate, rapid and easy to utilize. In addition, some platforms may work without using expensive equipment, because methods for fast and simple sample preparation have already been developed, whereas modern preamplification approaches allow to avoid applying thermocycling devices. Interestingly, a great amount of diverse types of natural CRISPR/Cas systems have been already discovered. Such abundance promotes development of multiple artificial systems, each of which exerting own unique characteristics. Therefore, a variety of diagnostic platforms with different properties are created on their basis that allows researchers and physicians to choose an optimal approach for performing specific tasks. For this reason, insights into structure and operation of CRISPR/Cas systems are necessary for selecting a suitable platform. The current classification of systems is based on such principles serving as the basis, in turn, for convenient evaluation of the very variety of molecular diagnostics platforms and presentation of the typical technical characteristics and nuances for each method. Thus, this review, which is mainly devoted to the platforms for molecular diagnostics of infectious diseases, also touches upon the issues of functioning, devices, and classification of CRISPR/Cas systems.

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CRISPR as a Diagnostic Tool

Cited by 48 publications

References 56 publications

Sensitive and Rapid Detection of Citrus Scab Using an RPA-CRISPR/Cas12a System Combined with a Lateral Flow Assay

Sensitive and Rapid Detection of Citrus Scab Using an RPA-CRISPR/Cas12a System Combined with a Lateral Flow Assay

CRISPR/Cas: A New Tool in the Research of Telomeres and Telomerase as Well as a Novel Form of Cancer Therapy

CRISPR/Cas-based diagnostic platforms

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