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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...

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Updated: Jan 18, 2026

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

EASY-edit: a toolbox for high-throughput single-step custom genetic editing in bacteria.

Maxence Lejars¹, Tomoya Maeda², Maude Guillier¹

¹Expression génétique microbienne, UMR8261 CNRS, Université Paris Cité, Institut de Biologie Physico-Chimique, Paris 75005, France.

Nucleic Acids Research

|September 10, 2025

Summary

We developed EASY-edit, a fast and affordable method for targeted gene editing in Escherichia coli. This engineered system simplifies the creation of genetic tools like mutant libraries and reporter genes, accelerating biological research and bioengineering.

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Published on: April 11, 2019

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Area of Science:

Molecular Biology
Synthetic Biology
Microbiology

Background:

CRISPR-Cas9 recombineering enables targeted gene editing but requires locus-specific optimization.
This optimization process can be inefficient, time-consuming, and labor-intensive.

Purpose of the Study:

To develop a simple, fast, and cost-effective method for high-efficiency gene editing in Escherichia coli.
To create a modular and flexible platform for constructing genetic tools and studying biological processes.

Main Methods:

Engineered Assembly of SYnthetic operons for targeted editing (EASY-edit) system was developed.
Utilized optimized guide RNAs and DNA repair templates with short homology arms.
Demonstrated modularity through iterative editing and combination of operon versions in three loci.

Main Results:

Achieved highly efficient editing of operon elements in Escherichia coli.
Successfully constructed reporter gene fusions validating post-transcriptional regulation.
Generated saturated and unbiased mutant libraries for genetic studies.

Conclusions:

The EASY-edit system offers a flexible genomic expression platform for biological research.
This method accelerates the development of genetic tools for bioengineering applications.
EASY-edit simplifies complex gene editing tasks, making them more accessible.