An aM-level sensitive cascade CRISPR-Dx system (ASCas) for rapid detection of RNA without pre-amplification

Zhang, Yibin; Chen, Yong; Zhang, Qianling; Liu, Yizhen; Zhang, Xueji

doi:10.1016/j.bios.2023.115248

Cited by 17 publications

(15 citation statements)

References 29 publications

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“…In contrast to RT‐qPCR, which requires an expensive fluorescent PCR instrument and relatively costly fluorescent probes, 7 RT‐PCR only necessitates a standard PCR instrument, while RT‐LAMP operates under isothermal conditions. Notably, some researchers have developed CRISPR cascade signal amplification assays that eliminate the need for pre‐amplification, 90 thus obviating the requirement for a PCR instrument and further reducing equipment costs. Second, CRISPR signal detection can be coupled with lateral flow assay technology, eliminating the need for specialized personnel and stringent experimental conditions 48 .…”

Section: Detection Of Vocs Using Crispr Systemmentioning

confidence: 99%

CRISPR‐Cas system: A promising tool for rapid detection of SARS‐CoV‐2 variants

Li,

Zhang,

Lin

et al. 2024

Journal of Medical Virology

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COVID‐19, caused by SARS‐CoV‐2, remains a global health crisis. The emergence of multiple variants with enhanced characteristics necessitates their detection and monitoring. Genome sequencing, the gold standard, faces implementation challenges due to complexity, cost, and limited throughput. The CRISPR‐Cas system offers promising potential for rapid variant detection, with advantages such as speed, sensitivity, specificity, and programmability. This review provides an in‐depth examination of the applications of CRISPR‐Cas in mutation detection specifically for SARS‐CoV‐2. It begins by introducing SARS‐CoV‐2 and existing variant detection platforms. The principles of the CRISPR‐Cas system are then clarified, followed by an exploration of three CRISPR‐Cas‐based mutation detection platforms, which are evaluated from different perspectives. The review discusses strategies for mutation site selection and the utilization of CRISPR‐Cas, offering valuable insights for the development of mutation detection methods. Furthermore, a critical analysis of the clinical applications, advantages, disadvantages, challenges, and prospects of the CRISPR‐Cas system is provided.

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Section: Detection Of Vocs Using Crispr Systemmentioning

confidence: 99%

CRISPR‐Cas system: A promising tool for rapid detection of SARS‐CoV‐2 variants

Li,

Zhang,

Lin

et al. 2024

Journal of Medical Virology

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“…230 , Zhang, Y.; Chen, Y.; Zhang, Q.; Liu, Y.; Zhang, X. An aM-level sensitive cascade CRISPR-Dx system (ASCas) for rapid detection of RNA without preamplification, 115248 (ref ). Copyright 2023, with permission from Elsevier.…”

Section: Crispr-based Direct Target Detectionmentioning

confidence: 99%

“…Additionally, utilizing a Cas13a-Cas12a cascade and three crRNAs, Zhang’s group developed an amplification-free, ultrasensitive, and ultrafast RNA detection approach termed ASCas (Figure D). This approach can detect SARS-CoV-2 at a concentration of 1 aM in just 20 min …”

Section: Crispr-based Direct Target Detectionmentioning

confidence: 99%

“…This, in turn, guides the “empty” Cas12a to recognize the assistant probe and cleave more ssDNA reporters to trigger the positive feedback circuit. In line with the rationale of CONAN, Zhang et al developed an ASCas system that combined a Cas13a-Cas12a cascade with multiple crRNAs, which realized the detection of SARS-CoV-2 with the unprecedented sensitivity (1 aM) of amplification-free methods …”

Section: Recycling-based Strategies For Target Amplification-free Dia...mentioning

confidence: 99%

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CRISPR-Powered Strategies for Amplification-Free Diagnostics of Infectious Diseases

Xia,

Rao,

Xiong

et al. 2024

Anal. Chem.

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“…Amplification-free detection of nucleic acids by CRISPR/Cas sensors requires a combination with other techniques and approaches such as SERS 8 , metal-enhanced fluorescence 9 , nanoenzymes 10 , and field-effect transistors 11 which require specialised instrumentation or consumables. The Cas tandem biosensing systems such as Cas13 with Csm6 tandem system 12 , Cas13 with Cas14 tandem system 13 , and Cas13 with Cas12 tandem system 14 offer a strategy for disrupting the one-to-one correspondence between target molecules and activated RNPs to realize one-to-more correspondence. However, all the current tandem systems are designed for the detection of RNA targets.…”

Section: Introductionmentioning

confidence: 99%

Hairpin-locker mediated CRISPR/Cas tandem system for ultrasensitive detection of DNA without pre-amplification

Deng,

Sang,

et al. 2024

Preprint

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Achieving ultra-sensitive detection of DNA is of paramount importance in the field of molecular analytics. Conventional amplification technologies such as polymerase chain reaction (PCR) currently play a leading role in ultrasensitive DNA detection. However, amplicon contamination common in these techniques may lead to false positives. To date, CRISPR-associated nucleases (type V & VI) with their programmable cleavage have been utilised for sensitive detection of unamplified nucleic acids in complex real samples. Nevertheless, without additional amplification strategies, the pM range sensitivity of such CRISPR/Cas sensors is not sufficient for clinical applications. Here, we established a hairpin-locker (H-locker) mediated Cas12-Cas13 tandem biosensing system (Cas12-13 tandem-sensor) for ultrasensitive detection of DNA targets. Without the need for any additional amplification reaction or device, this system is capable of detecting DNA at a notable 1 aM level (<1 copy/uL) sensitivity. In addition, the system was able to distinguish cancer mutations in colorectal cancer (CRC) mice. This is a significant advance for CRISPR/Cas biosensing technology offering simple, highly sensitive, and user-friendly diagnostics for next-generation nucleic acid detection.

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An aM-level sensitive cascade CRISPR-Dx system (ASCas) for rapid detection of RNA without pre-amplification

Cited by 17 publications

References 29 publications

CRISPR‐Cas system: A promising tool for rapid detection of SARS‐CoV‐2 variants

CRISPR‐Cas system: A promising tool for rapid detection of SARS‐CoV‐2 variants

CRISPR-Powered Strategies for Amplification-Free Diagnostics of Infectious Diseases

Hairpin-locker mediated CRISPR/Cas tandem system for ultrasensitive detection of DNA without pre-amplification

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