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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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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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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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Updated: Nov 6, 2025

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
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CRISPR Enzyme Kinetics for Molecular Diagnostics.

Ashwin Ramachandran1, Juan G Santiago1

  • 1Department of Mechanical Engineering, Stanford University, Stanford, California 94305, United States.

Analytical Chemistry
|May 12, 2021
PubMed
Summary
This summary is machine-generated.

CRISPR enzyme kinetics were modeled using Michaelis-Menten theory, revealing widespread violations of physical limits in published data. This work informs achievable detection limits and assay times for CRISPR-based diagnostics.

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Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • CRISPR-diagnostic assays have surged in interest, particularly for rapid testing during the COVID-19 pandemic.
  • Understanding the kinetic limits of CRISPR enzymes like Cas12 and Cas13 is crucial for optimizing diagnostic performance.

Purpose of the Study:

  • To develop a model based on Michaelis-Menten enzyme kinetics to analyze CRISPR enzyme activity.
  • To establish criteria for validating reported kinetic parameters and assess their physical consistency.
  • To explore the implications of enzyme kinetics on achievable limits of detection and assay times for CRISPR diagnostics.

Main Methods:

  • Applied Michaelis-Menten enzyme kinetics theory to model CRISPR enzymes (Cas12, Cas13).
  • Developed analytical solutions for reaction kinetics and validation criteria for kinetic parameters.
  • Analyzed published kinetic data for various Cas12 and Cas13 subtypes and orthologs.
  • Conducted experimental studies on LbCas12a kinetics with ssDNA and dsDNA activators to validate the model.

Main Results:

  • A significant majority of previously reported CRISPR enzyme kinetic data violate basic physical limits.
  • The developed model, validated by experimental data, accurately predicts CRISPR reaction kinetics.
  • Demonstrated that enzyme kinetics significantly impact achievable limits of detection and assay times for CRISPR-based diagnostic assays.

Conclusions:

  • Published kinetic data for many CRISPR enzymes require re-evaluation due to apparent violations of physical principles.
  • The validated kinetic model provides a framework for predicting and optimizing the performance of CRISPR-diagnostic assays.
  • This research has broad implications for the development of more sensitive and rapid amplification-free CRISPR-detection methods.