Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a

Lu, Shuhan; Tong, Xiaohan; Han, Yang; Zhang, Kun; Zhang, Yizhou; Chen, Qiubing; Duan, Junyi; Lei, Xinlin; Huang, Mingbin; Qiu, Yang; Zhang, Ding-Yu; Zhou, Xi; Zhang, Ying; Yin, Hao

doi:10.1038/s41551-022-00861-x

Cited by 149 publications

(133 citation statements)

References 39 publications

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“…Diagnostic CRISPR systems have outstanding superiority in terms of their ultrahigh sensitivity, portability, and specificity. For instance, the Cas12 and Cas13 nucleases have been reprogrammed to detect the nucleic acids of SARS‐CoV‐2 134–136 …”

Section: Genome Editing For Fundamental Research: Disease Modelling A...mentioning

confidence: 99%

“…For instance, the Cas12 and Cas13 nucleases have been reprogrammed to detect the nucleic acids of SARS-CoV-2. [134][135][136] Viruses, whether DNA viruses or RNA viruses, are important pathogens of infectious diseases. RNA viruses that have been studied abundantly in recent years include dengue, 137 Zika, 49 and a recent spotlight of interest, SARS-CoV-2.…”

Section: New Disease Diagnostic Tools Based On the Crispr-cas Systemmentioning

confidence: 99%

“…140,141 In 2022, Lu et al reduced the detection time of CRISPR-based assays while ensuring accuracy through a hybrid strategy, which includes adjusting the kinetics of Cas12a and using more flexible crRNA designs. 134 In addition to RNA viruses, CRISPR/Cas-based diagnostic methods can identify DNA viruses, such as BK virus (BKV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). 142,143 The researchers used the SHERLOCK system to detect BKV or CMV in serum samples and then verified the results by qPCR.…”

Section: New Disease Diagnostic Tools Based On the Crispr-cas Systemmentioning

confidence: 99%

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Current landscape of gene‐editing technology in biomedicine: Applications, advantages, challenges, and perspectives

Zhou

Yang

Zhang

et al. 2022

MedComm

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The expanding genome editing toolbox has revolutionized life science research ranging from the bench to the bedside. These “molecular scissors” have offered us unprecedented abilities to manipulate nucleic acid sequences precisely in living cells from diverse species. Continued advances in genome editing exponentially broaden our knowledge of human genetics, epigenetics, molecular biology, and pathology. Currently, gene editing‐mediated therapies have led to impressive responses in patients with hematological diseases, including sickle cell disease and thalassemia. With the discovery of more efficient, precise and sophisticated gene‐editing tools, more therapeutic gene‐editing approaches will enter the clinic to treat various diseases, such as acquired immunodeficiency sydrome (AIDS), hematologic malignancies, and even severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection. These initial successes have spurred the further innovation and development of gene‐editing technology. In this review, we will introduce the architecture and mechanism of the current gene‐editing tools, including clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR‐associated nuclease‐based tools and other protein‐based DNA targeting systems, and we summarize the meaningful applications of diverse technologies in preclinical studies, focusing on the establishment of disease models and diagnostic techniques. Finally, we provide a comprehensive overview of clinical information using gene‐editing therapeutics for treating various human diseases and emphasize the opportunities and challenges.

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Section: Genome Editing For Fundamental Research: Disease Modelling A...mentioning

confidence: 99%

Section: New Disease Diagnostic Tools Based On the Crispr-cas Systemmentioning

confidence: 99%

Section: New Disease Diagnostic Tools Based On the Crispr-cas Systemmentioning

confidence: 99%

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Current landscape of gene‐editing technology in biomedicine: Applications, advantages, challenges, and perspectives

Zhou

Yang

Zhang

et al. 2022

MedComm

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“…It was recently demonstrated that it is possible to combine isothermal amplification and CRISPR detection in a one-pot reaction via delicately engineering the primer designs and reaction conditions. 27,[34][35][36] However, it remains unclear if such strategy can be adapted to develop one-step, one-pot CRISPR assays for miRNA sensing. Alternatively, CRISPRmediated target recognition can be integrated with other signal transduction modalities, such as electrochemical, 37 plasmonic, 38 and graphene field-effect transistor (gFET) sensors, 39 enabling amplification-free nucleic acid detection.…”

Section: Introductionmentioning

confidence: 99%

One-Pot Endonucleolytically Exponentiated Rolling Circle Amplification by CRISPR-Cas12a Affords Sensitive, Expedited Isothermal Detection of MicroRNAs

Wen

Han

et al. 2022

Preprint

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MicroRNAs (miRNAs) are a class of short non-coding RNAs that play essential roles in gene expression regulation. While miRNAs offer a promising source for developing potent cancer biomarkers, the progress towards clinical utilities remains largely limited, due in part to the long-standing challenge in sensitive, specific, and robust detection of miRNAs in human biofluids. Emerging next-generation molecular technologies, such as the CRISPR-based methods, promise to transform nucleic acid testing. The prevailing strategy used in existing CRISPR-based methods is to hyphenate two separate reactions for pre-amplification, e.g., rolling circle amplification (RCA), and amplicon detection by Cas12a/13a trans-cleavage in tandem. Thus, existing CRISPR-based miRNA assays require multiple manual steps and lack the analytical performance of the gold standard, RT-qPCR. Radically deviating from the existing strategies, we developed a one-step, one-pot isothermal miRNA assay termed “Endonucleolytically eXponenTiated Rolling circle Amplification with the dual-functional CRISPR-Cas12a” (EXTRA-CRISPR) to afford RT-PCR-like performance for miRNA detection. We demonstrated the superior analytical performance of our EXTRA-CRISPR assay to detect miRNAs (miR-21, miR-196a, miR-451a, and miR-1246) in plasma extracellular vesicles, which allowed us to define a potent EV miRNA signature for detection of pancreatic cancer. The analytical and diagnostic performance of our one-pot assay were shown to be comparable with that of the commercial RT-qPCR assays, while greatly simplifying and expediting the analysis workflow. Therefore, we envision that our technology provides a promising tool to advance miRNA analysis and clinical marker development for liquid biopsy-based cancer diagnosis and prognosis.

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“…Since December 2019, the coronavirus disease 2019 (COVID-19), caused by a novel coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been continuously spreading and rapidly becoming a global pandemic as declared by the World Health Organization . Although the specific vaccination coverage has been rising rapidly around the world, recent research has reported that the protection rate of vaccination could hardly reach 100%. − Hence, rapid and accurate population screening is still crucial for the control of COVID-19 spreading, , especially at airports or hospitals where large-scale screening is required. Optimal detection methods are expected to be on-site, rapid, simple, safe, and portable. , …”

Section: Introductionmentioning

confidence: 99%

Integrated System for On-Site Rapid and Safe Screening of COVID-19

et al. 2022

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Since the outbreak of coronavirus disease 2019 (COVID-19), the epidemic has been spreading around the world for more than 2 years. Rapid, safe, and on-site detection methods of COVID-19 are in urgent demand for the control of the epidemic. Here, we established an integrated system, which incorporates a machine-learning-based Fourier transform infrared spectroscopy technique for rapid COVID-19 screening and air-plasma-based disinfection modules to prevent potential secondary infections. A partial least-squares discrimination analysis and a convolutional neural network model were built using the collected infrared spectral dataset containing 857 training serum samples. Furthermore, the sensitivity, specificity, and prediction accuracy could all reach over 94% from the results of the field test regarding 968 blind testing samples. Additionally, the disinfection modules achieved an inactivation efficiency of 99.9% for surface and airborne tested bacteria. The proposed system is conducive and promising for point-of-care and on-site COVID-19 screening in the mass population.

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Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a

Cited by 149 publications

References 39 publications

Current landscape of gene‐editing technology in biomedicine: Applications, advantages, challenges, and perspectives

Current landscape of gene‐editing technology in biomedicine: Applications, advantages, challenges, and perspectives

One-Pot Endonucleolytically Exponentiated Rolling Circle Amplification by CRISPR-Cas12a Affords Sensitive, Expedited Isothermal Detection of MicroRNAs

Integrated System for On-Site Rapid and Safe Screening of COVID-19

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