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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...

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Related Experiment Video

Updated: May 2, 2026

CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus AAV and 2-Cell Embryo Electroporation

CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus AAV and 2-Cell Embryo Electroporation

Published on: March 15, 2024

Heritable multiplex genetic engineering in rats using CRISPR/Cas9.

Yuanwu Ma¹, Bin Shen², Xu Zhang¹

¹Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Beijing, China.

|March 7, 2014

Summary

This summary is machine-generated.

The CRISPR/Cas9 gene-editing tool efficiently disrupts four genes simultaneously in rats. This method reliably generates germline transmissible gene knock-out rats with rare off-target mutations.

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

The CRISPR/Cas9 system is a powerful tool for genome modification.
Multiplex genetic engineering in rats is crucial for studying complex diseases.

Purpose of the Study:

To demonstrate the efficiency of CRISPR/Cas9 for simultaneous disruption of four genes in rats.
To confirm germline transmission and assess off-target effects of the gene editing.

Main Methods:

Co-injection of Cas9 mRNA and sgRNAs targeting four genes (ApoE, B2m, Prf1, Prkdc) into fertilized rat eggs.
Analysis of gene modifications, germline transmissibility, off-target mutagenesis, and mosaicism.

Main Results:

Simultaneous disruption of four target genes was achieved in a single step.
Gene modifications were confirmed to be germline transmittable.
Off-target mutagenesis and mosaicism were detected at very low frequencies.

Conclusions:

The CRISPR/Cas9 system provides an efficient and reliable method for generating multi-gene knock-out rats.
This technique facilitates the study of complex genetic diseases using rat models.