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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/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 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...

Homologous Recombination

Homologous Recombination

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...

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Related Experiment Video

Updated: Dec 29, 2025

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Published on: June 2, 2018

CRISPR/Cas9 Guide RNA Design Rules for Predicting Activity.

Kasidet Hiranniramol¹, Yuhao Chen¹, Xiaowei Wang²

¹Department of Radiation Oncology, Washington University School of Medicine, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)

|February 2, 2020

Summary

This summary is machine-generated.

Designing effective guide RNA (gRNA) is crucial for CRISPR/Cas9 gene editing success. This review covers gRNA design rules to enhance on-target activity and reduce off-target effects, alongside computational tools for prediction.

Keywords:

CRISPR Cas9 Genome editing Machine learning sgRNA

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Last Updated: Dec 29, 2025

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

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CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

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Published on: April 25, 2022

Area of Science:

Molecular Biology
Genetics
Bioinformatics

Background:

The CRISPR/Cas9 system is a powerful tool for gene editing.
Effective guide RNA (gRNA) design is essential for successful CRISPR/Cas9 experiments.
Maximizing on-target activity and minimizing off-target effects are key challenges in gRNA design.

Purpose of the Study:

To review current gRNA design rules for CRISPR/Cas9.
To identify strategies for maximizing on-target Cas9 activity.
To outline methods for minimizing off-target Cas9 activity.

Main Methods:

Literature review of existing gRNA design principles.
Analysis of computational tools for gRNA design and prediction.
Evaluation of assays for gRNA activity assessment.

Main Results:

Established gRNA design rules that correlate with high on-target activity.
Identified factors contributing to off-target Cas9 activity.
Cataloged available computational tools for gRNA design and prediction.

Conclusions:

Optimized gRNA design is critical for efficient and specific gene editing.
Computational tools can aid in selecting effective gRNAs.
Further development of design rules and prediction tools will enhance CRISPR/Cas9 applications.