Related Experiment Video
Updated: Dec 27, 2025
08:23
CIRCLE-Seq for Interrogation of Off-Target Gene Editing
Published on: November 1, 2024
1.3K
CRISPR system: Discovery, development and off-target detection.
Shengmiao Chen1, Yufeng Yao1, Yanchun Zhang1
1School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
Cellular Signalling
|February 26, 2020
Summary
The CRISPR-Cas system offers revolutionary gene editing but faces off-target effects. This review summarizes breakthroughs and methods for detecting these effects to improve genomic editing accuracy.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- CRISPR-Cas is a powerful gene editing tool derived from microbial adaptive immunity.
- Compared to ZFNs and TALENs, CRISPR-Cas offers significant advantages.
- Off-target mutations are a key limitation hindering clinical applications of CRISPR-Cas.
Purpose of the Study:
- To review major advancements in CRISPR-Cas system development and application.
- To summarize current methods for detecting off-target effects in CRISPR-Cas mediated genetic manipulation.
- To aid researchers in achieving accurate genomic editing.
Main Methods:
- Literature review of CRISPR-Cas system development.
- Analysis of studies focusing on off-target effect detection methods.
- Synthesis of information on CRISPR-Cas applications and limitations.
Main Results:
- CRISPR-Cas technology has seen rapid development and diverse applications.
- Various methods exist for identifying and quantifying CRISPR-Cas off-target effects.
- Understanding and mitigating off-target effects are crucial for reliable gene editing.
Conclusions:
- CRISPR-Cas is a transformative gene editing technology with ongoing improvements.
- Accurate detection of off-target effects is essential for safe and effective translational research.
- This review provides a resource for scientists to enhance precision in genomic editing.
Related Concept Videos
01:59CRISPR
57.3K
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...
57.3K
01:28CRISPR/Cas9 Genome Editing
1.5K
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...
1.5K
02:53CRISPR and crRNAs
18.6K
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...
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...
18.6K
01:23The Antiviral System of Bacteria and Archaea: CRISPR
537
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
537
02:31Homologous Recombination
62.0K
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...
62.0K
02:53Conservative Site-specific Recombination and Phase Variation
6.5K
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.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.5K