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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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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.
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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Related Experiment Video

Updated: Jan 8, 2026

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
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A Versatile CRISPR/Cas12a Autocatalytic Cascade System via Structure-Switching V-Type Split Probe for Highly

Zhun Lin1, Zhe Pu1, Jiacheng Wu1

  • 1State Key Laboratory of Anti-Infective Drug Discovery and Development; School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China.

Analytical Chemistry
|December 15, 2025
PubMed
Summary

This study introduces a new, amplification-free CRISPR/Cas diagnostic system for rapid pathogen nucleic acid detection. It achieves atto-molar sensitivity, offering a simpler and faster alternative for infectious disease diagnostics.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Infectious Disease Diagnostics

Background:

  • Accurate and rapid detection of pathogen nucleic acids is crucial for managing infectious diseases.
  • Current molecular diagnostics, including CRISPR/Cas systems, often require preamplification, leading to complex workflows and expensive equipment.

Purpose of the Study:

  • To develop a rapid, simple, and amplification-free CRISPR/Cas-based diagnostic system.
  • To enhance sensitivity and reduce complexity in pathogen detection methods.

Main Methods:

  • Utilized a structure-switching V-shaped DNA probe with a split Cas12a recognition sequence.
  • Implemented a positive feedback loop and signal amplification cascade for exponential signal generation.
  • Integrated the system into microfluidic and lateral flow assays.

Main Results:

  • Achieved an ultralow background signal with rapid, exponential signal production.
  • Demonstrated atto-molar sensitivity for pathogen DNA detection.
  • Successfully applied to multiplex detection of human papillomavirus strains and point-of-care detection of monkeypox virus.

Conclusions:

  • The developed amplification-free CRISPR/Cas system offers a significant advancement in rapid and sensitive pathogen DNA detection.
  • This approach holds great potential for both clinical laboratory and point-of-care diagnostic applications.
  • Simplifies workflows and reduces reliance on expensive equipment for infectious disease diagnostics.