The CRISPR–Cas toolbox for analytical and diagnostic assay development

Tang, Yanan; L, Gao; Feng, Wei; Guo, Chuanbin; Yang, Qianfan; Li, Feng; Le, X. Chris

doi:10.1039/d1cs00098e

Cited by 125 publications

(109 citation statements)

References 190 publications

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“…Recently, CRISPR-Cas has emerged as a powerful technology that is revolutionizing next-generation diagnostic platforms [6] , [7] . The mechanism of CRISPR-Cas systems, which are programmed by CRISPR guide RNA (crRNA), has been reported to not only cleave the target nucleic acid but also cleave all neighbouring nucleic acids indiscriminately [8] , [9] .…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection

Liu

Zhang

2022

Bioelectrochemistry

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Section: Introductionmentioning

confidence: 99%

CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection

Liu

Zhang

2022

Bioelectrochemistry

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“…The emergence of the CRISPR/Cas system produced a revolution in gene editing technology. CRISPR/Cas is an acquired immune system in bacteria that is used to fight invading DNA, plasmids, and phages [ 11 ]. The CRISPR/Cas system consists of CRISPR-derived RNA (crRNA) and Cas proteins.…”

Section: Gene Editing Technologiesmentioning

confidence: 99%

Reprogramming the tumor microenvironment by genome editing for precision cancer therapy

et al. 2022

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The tumor microenvironment (TME) is essential for immune escape by tumor cells. It plays essential roles in tumor development and metastasis. The clinical outcomes of tumors are often closely related to individual differences in the patient TME. Therefore, reprogramming TME cells and their intercellular communication is an attractive and promising strategy for cancer therapy. TME cells consist of immune and nonimmune cells. These cells need to be manipulated precisely and safely to improve cancer therapy. Furthermore, it is encouraging that this field has rapidly developed in recent years with the advent and development of gene editing technologies. In this review, we briefly introduce gene editing technologies and systematically summarize their applications in the TME for precision cancer therapy, including the reprogramming of TME cells and their intercellular communication. TME cell reprogramming can regulate cell differentiation, proliferation, and function. Moreover, reprogramming the intercellular communication of TME cells can optimize immune infiltration and the specific recognition of tumor cells by immune cells. Thus, gene editing will pave the way for further breakthroughs in precision cancer therapy.

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“…For example, CRISPR diagnostics are viewed as a serious contender for fast and deployable assays for the detection of the RNA of SARS-CoV-2, the virus that causes Covid-19 (Ramachandran et al 2020;Fozouni et al 2021;Shinoda et al 2021;Nguyen, Smith, and Jain 2020). The most commonly applied Cas types for https://doi.org/10.1017/qrd.2022.7 Published online by Cambridge University Press diagnostics are Cas12 and Cas13, which offer direct detection of DNA and RNA targets, respectively (Tang et al 2021).…”

Section: Introductionmentioning

confidence: 99%

“…As a diagnostic tool, CRISPR-Cas systems offer distinct advantages as they can be reconfigured using synthetic guide RNAs; are compatible with a few detection modalities including fluorescent reporter probes; are compatible with lyophilized reagents; and will likely be field-deployable, particularly given robust enzyme activity at ambient temperatures (Tang et al 2021). This potential has also led to significant investments in new biotechnology companies pursuing CRISPR diagnostic as products, notably Mammoth Biosciences, Inc. in Brisbane, California and Sherlock Biosciences in Cambridge, Massachusetts.…”

Section: Introductionmentioning

confidence: 99%

Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential

Santiago

2022

QRB Discovery

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The scientific and technological advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is one of the most exciting developments of the past decade, particularly in the field of gene editing. The technology has two essential components, (1) a guide RNA to match a targeted gene, and (2) a CRISPR-associated protein (e.g., Cas 9, Cas12, or Cas13) that acts as an endonuclease to specifically cut DNA. This specificity and reconfigurable nature of CRISPR has also spurred intense academic and commercial interest in the development of CRISPR-based molecular diagnostics. CRISPR Cas12 and Cas13 orthologs are most commonly applied to diagnostics, and these cleave and become activated by DNA and RNA targets, respectively. Despite the resulting intense research activity, the possible limits of detection and applications of CRISP-based diagnostics remain an open question. A major reason for this is that reports of CRISPR-Cas kinetic rates have been widely inconsistent, and the vast majority of these reports have been shown to contain gross errors including violations of basic conservation and kinetic rate laws. It is the intent of this Perspective to bring attention to these issues and to, more importantly, identify potential improvements in the manner in which CRISPR kinetic rates and assay LoDs are reported and compared. The CRISPR field would benefit from formal checks of self-consistency of data, providing sufficient information and raw data such that experiments can be reproduced, and, in the case of publications that report LoDs of novel assays, concurrent and clear reporting of the kinetic rate constants responsible for such LoDs. The early development of CRISPR-based diagnostics calls for self-reflection and urges all of us to proceed with caution.

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The CRISPR–Cas toolbox for analytical and diagnostic assay development

Abstract: A comprehensive review that offers mechanistic insight into the CRISPR–Cas toolbox for analytical and diagnostic assay development.

Cited by 125 publications

References 190 publications

CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection

CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection

Reprogramming the tumor microenvironment by genome editing for precision cancer therapy

Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential

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