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Related Concept Videos

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CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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The Antiviral System of Bacteria and Archaea: CRISPR01:23

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Related Experiment Video

Updated: Sep 3, 2025

DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
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DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning

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Development of efficient, sensitive, and specific detection method for Encephalomyocarditis virus based on

Ning Wei1, Junyao Xiong1, Junheng Ma1

  • 1State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.

Journal of Virological Methods
|July 29, 2022
PubMed
Summary

A new CRISPR/Cas13a-based method offers rapid, sensitive, and specific detection of Encephalomyocarditis virus (EMCV). This isothermal assay, combining recombinase-aided amplification and lateral flow, provides accurate results within an hour for improved disease diagnosis and control.

Keywords:
CRISPR/Cas13aDetection methodEncephalomyocarditis virusLateral flow stripRAA

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Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Encephalomyocarditis virus (EMCV) is a significant zoonotic pathogen causing substantial economic losses in swine.
  • Accurate and rapid detection of EMCV is crucial for disease diagnosis and control.
  • CRISPR/Cas13a systems offer precise RNA targeting with collateral cleavage activity.

Purpose of the Study:

  • To develop an efficient, sensitive, and specific detection method for EMCV.
  • To leverage the collateral cleavage activity of CRISPR/Cas13a for viral detection.
  • To create a rapid, isothermal diagnostic assay for EMCV.

Main Methods:

  • Combined recombinase-aided amplification (RAA) with CRISPR/Cas13a and a lateral flow strip for EMCV detection.
  • Utilized the collateral RNase activity of LwCas13a upon target recognition.
  • Optimized isothermal reaction conditions for detection at 37°C.

Main Results:

  • Achieved a detection sensitivity of up to 10^1 copies/µL for EMCV.
  • Demonstrated excellent specificity with no cross-reactivity against 8 other major swine viruses.
  • Showcased a 100% coincidence rate with qPCR in 37 clinical samples.
  • Obtained results within 60 minutes with visual observation.

Conclusions:

  • The developed CRISPR/Cas13a-based method is a simple, inexpensive, and highly effective tool for EMCV detection.
  • This assay has significant potential for rapid and accurate diagnosis of EMCV in clinical settings.
  • The method's isothermal nature and visual readout facilitate its application in resource-limited environments.