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CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

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...

CRISPR

CRISPR

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...

CRISPR

CRISPR

CRISPR and crRNAs

CRISPR and crRNAs

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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Updated: Apr 20, 2026

Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes

Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes

Published on: May 23, 2025

Imaging genomic elements in living cells using CRISPR/Cas9.

Baohui Chen¹, Bo Huang²

¹Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA.

Methods in Enzymology

|November 16, 2014

Summary

This summary is machine-generated.

CRISPR and TALE systems can now visualize DNA in living cells, aiding genome function studies. Protocols detail using CRISPR/Cas9 to label and image specific genomic loci, including repetitive and non-repetitive sequences.

Keywords:

CRISPR Chromatin Fluorescence Microscopy Nucleus Telomere

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CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

Area of Science:

Molecular Biology
Genomics
Cell Biology

Background:

Programmable DNA recognition systems like CRISPR and TALE are crucial for genome editing and gene regulation.
These systems are now engineered for visualizing endogenous genomic elements in living cells.
Understanding genome organization and nuclear interactions is key to studying gene function regulation.

Purpose of the Study:

To discuss general considerations for designing and implementing DNA imaging systems in living cells.
To provide detailed protocols for using the CRISPR/Cas9 system to label and image specific genomic loci.
To demonstrate the application of these systems for studying genome function through physical organization.

Main Methods:

Establishing expression systems for catalytically inactive Cas9 fused to GFP (dCas9-GFP) and single-guide RNA (sgRNA).
Developing procedures for labeling repetitive genomic sequences, such as telomeres and protein-coding genes.
Implementing simultaneous expression of multiple sgRNAs for labeling non-repetitive genomic loci.
Utilizing Fluorescence In Situ Hybridization (FISH) for signal specificity verification.

Main Results:

Successful labeling and imaging of specific genomic loci in living cells using CRISPR/Cas9.
Demonstrated ability to target both repetitive and non-repetitive genomic regions.
Verified the specificity of the labeling system through FISH analysis.

Conclusions:

CRISPR/Cas9-based systems offer a powerful tool for visualizing endogenous genomic elements in living cells.
These imaging capabilities significantly advance the study of genome function by revealing physical organization and interactions.
The provided protocols enable researchers to implement and validate DNA labeling strategies for diverse genomic targets.