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

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

2.1K
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...
2.1K
CRISPR01:59

CRISPR

58.2K
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...
58.2K
CRISPR and crRNAs02:53

CRISPR and crRNAs

19.3K
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...
19.3K

You might also read

Related Articles

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

Sort by
Same author

CAR T cell therapy selectively depletes disease-driving mutant calreticulin cells in xenotransplants and human organoid models of myelofibrosis.

Science translational medicine·2026
Same author

Nanobodies to GPVI as alternative reagents for platelet spreading.

Platelets·2026
Same author

Engineering bone marrow in a dish-a bloody business: preclinical opportunities, translational use cases, and a call for consensus.

Journal of thrombosis and haemostasis : JTH·2026
Same author

comBO: A combined human bone and lympho-myeloid bone marrow organoid for preclinical modeling of hematopoietic disorders.

Cell stem cell·2026
Same author

Formation of tight junction-like structures of zonula occludens 2 in platelet-platelet interaction.

Research and practice in thrombosis and haemostasis·2026
Same author

Human Multi-Organoid Platform to Model Immune Dynamics in Cardiac Injury and Disease.

Circulation research·2025

Related Experiment Video

Updated: Feb 24, 2026

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish
09:38

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish

Published on: January 24, 2025

1.3K

CRISPR-Cas9 Mediated Labelling Allows for Single Molecule Imaging and Resolution.

Abdullah O Khan1, Victoria A Simms1, Jeremy A Pike2

  • 1Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK.

Scientific Reports
|August 18, 2017
PubMed
Summary

CRISPR-Cas9 gene editing enables high-resolution live cell imaging using photoactivated localization microscopy (PALM) at endogenous protein levels. This method achieves nanoscopic resolution comparable to dSTORM, overcoming previous limitations in quantitative single-molecule super-resolution microscopy.

More Related Videos

Super-Resolution Microscopy of the Synaptonemal Complex Within the Caenorhabditis elegans Germline
09:14

Super-Resolution Microscopy of the Synaptonemal Complex Within the Caenorhabditis elegans Germline

Published on: September 13, 2022

3.1K
Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

12.1K

Related Experiment Videos

Last Updated: Feb 24, 2026

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish
09:38

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish

Published on: January 24, 2025

1.3K
Super-Resolution Microscopy of the Synaptonemal Complex Within the Caenorhabditis elegans Germline
09:14

Super-Resolution Microscopy of the Synaptonemal Complex Within the Caenorhabditis elegans Germline

Published on: September 13, 2022

3.1K
Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

12.1K

Area of Science:

  • Biophysics
  • Cell Biology
  • Molecular Imaging

Background:

  • Super-resolution microscopy techniques like dSTORM and PALM offer nanoscale insights into cellular structures.
  • Accurate quantification and spatial relationship analysis in single-molecule localization microscopy (SMLM) face challenges.
  • CRISPR-Cas9 gene editing provides a method for stable cell line generation with endogenous protein expression levels for molecular probes.

Purpose of the Study:

  • To investigate the feasibility of using CRISPR-Cas9 gene editing for photoactivated localization microscopy (PALM) probe generation.
  • To determine the relationship between endogenous expression levels and PALM imaging quality.
  • To assess if CRISPR-generated PALM probes can achieve resolutions comparable to established super-resolution methods.

Main Methods:

  • CRISPR-Cas9 gene editing was employed to tag an endogenous cytoskeletal protein (α-tubulin) with PALM fluorophores.
  • The expression levels of the tagged protein were analyzed in relation to imaging quality.
  • Photoactivated localization microscopy (PALM) was performed on the generated cell lines.

Main Results:

  • A direct correlation was established between protein expression level and the quality of PALM imaging.
  • CRISPR-PALM achieved spatial resolutions comparable to dSTORM.
  • The choice of tag influenced the total expression of the labeled protein, impacting imaging outcomes.

Conclusions:

  • CRISPR-PALM enables nanoscopic spatial resolution and quantitative benefits of SMLM via endogenous protein expression.
  • This approach facilitates super-resolved live-cell imaging with reduced biological variability.
  • Findings have implications for developing improved PALM tags and advancing super-resolution microscopy applications.