Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

CRISPR01:59

CRISPR

57.4K
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...
57.4K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.6K
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...
1.6K
CRISPR and crRNAs02:53

CRISPR and crRNAs

18.7K
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...
18.7K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A predictive model combining contrast-enhanced ultrasound and Shamblin classification for risk stratification of internal carotid artery resection in carotid body tumor surgery.

Quantitative imaging in medicine and surgery·2026
Same author

Label-free detection of microRNA by polymerization and isomerization cyclic amplification coupled with G/Hemin DNAzyme.

Journal of pharmaceutical analysis·2026
Same author

DNA-encoded library screening identifies CDK2-targeting lead compounds with favorable drug-like properties for anticancer development.

Journal of pharmaceutical analysis·2026
Same author

Potential for distinguishing the parkinsonian subtype of multiple system atrophy from Parkinson's disease: a three-dimensional gait analysis study.

Frontiers in aging neuroscience·2026
Same author

Controlling Enantioselectivity of Halohydrin Dehalogenase for Asymmetric Synthesis of Chiral Epichlorohydrin.

Biotechnology and bioengineering·2026
Same author

A Wearable Acetone Gas Sensor Enabled by Quantum Dot-Sensitized Flower-like Ti<sub>3</sub>C<sub>2</sub>T<sub><i>x</i></sub> for Metabolic Monitoring.

ACS sensors·2025

Related Experiment Video

Updated: Jan 9, 2026

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
10:16

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Published on: August 16, 2024

2.0K

Site accessibility-driven CRISPR/Cas13a activation for amplification-free RNA biosensing.

Zhaleh Asadi Fakhr1, Wei Xie1, Su Zeng1

  • 1State Key Laboratory of Advanced Drug Delivery and Release Systems, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China.

Analytica Chimica Acta
|December 6, 2025
PubMed
Summary

Site accessibility is key for CRISPR-Cas13a RNA diagnostics. Optimizing guide RNA (gRNA) structure improves enzyme activation, leading to more sensitive and rapid amplification-free RNA detection for point-of-care applications.

Keywords:
Atomic force microscopy (AFM)CRISPR-Cas13aEnzyme activation efficiencyIsothermal titration calorimetry (ITC)Michaelis-Menten kinetic modeling

More Related Videos

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
09:03

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

3.1K
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

2.2K

Related Experiment Videos

Last Updated: Jan 9, 2026

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
10:16

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Published on: August 16, 2024

2.0K
Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
09:03

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

3.1K
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

2.2K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Biophysics

Background:

  • CRISPR-Cas13a biosensing offers rapid, amplification-free RNA diagnostics.
  • Assay sensitivity is limited by guide RNA (gRNA) activation efficiency.
  • The roles of gRNA-target binding affinity and target site accessibility in activation remain unclear.

Purpose of the Study:

  • To systematically investigate the contributions of gRNA-target binding affinity and target site accessibility to CRISPR-Cas13a activation efficiency.
  • To establish clear design rules for optimizing gRNA selection in CRISPR-based diagnostics.

Main Methods:

  • Designed three ciRS-7-specific gRNAs with varying spacer accessibility (high, intermediate, low).
  • Quantified site accessibility and binding affinity using isothermal titration calorimetry (ITC).
  • Determined enzyme kinetics (kcat) using Michaelis-Menten analysis.
  • Evaluated assay sensitivity through detection-limit experiments.

Main Results:

  • Site accessibility directly correlated with catalytic turnover (kcat), with higher accessibility leading to greater efficiency.
  • The gRNA with higher site accessibility, despite lower binding affinity, showed superior Cas13a activation.
  • gRNAs with greater spacer accessibility demonstrated enhanced signal strength and improved detection limits.

Conclusions:

  • Spacer accessibility is a critical determinant of Cas13a enzyme activation in amplification-free RNA sensing.
  • Prioritizing unstructured spacer regions in gRNA design enhances enzyme activation efficiency.
  • This strategy provides a design rule for developing next-generation CRISPR diagnostics with improved speed and sensitivity.