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Updated: Oct 23, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Principles of Genetic Engineering.

Thomas M Lanigan^1,2, Huira C Kopera^1,3, Thomas L Saunders^4,5

¹Biomedical Research Core Facilities, Vector Core, University of Michigan, Ann Arbor, MI 48109, USA.

|March 14, 2020

Summary

This summary is machine-generated.

Genetic engineering uses molecular biology to modify DNA. CRISPR/Cas9 technology is now the leading method for gene targeting, surpassing older techniques due to its precision and efficiency in genome editing.

Keywords:

CRISPR/Cas9 embryonic stem (ES) cells gene targeting genetic engineering homologous recombination microinjection retroviruses transgenic mice transgenic rats transposons vectors

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Last Updated: Oct 23, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Generation of Genetically Modified Mice through the Microinjection of Oocytes

Generation of Genetically Modified Mice through the Microinjection of Oocytes

Published on: June 15, 2017

Mouse Genome Engineering Using Designer Nucleases

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Traditional genetic engineering methods like homologous recombination are inefficient and cumbersome.
Random DNA integration via transfection or viral vectors has drawbacks despite higher efficiency.
Guided endonucleases offer precise DNA targeting, revolutionizing genome modification.

Purpose of the Study:

To outline the fundamental principles of genetic engineering.
To detail current technologies and their common elements.
To discuss the evolution of gene targeting methods.

Main Methods:

Review of established genetic engineering techniques (homologous recombination, random integration).
Focus on endonuclease-based technologies, particularly CRISPR/Cas9.
Discussion of essential components: chromosome breaks, genotyping assays, and delivery methods.

Main Results:

CRISPR/Cas9 technology has become the dominant method for gene targeting.
Endonuclease-mediated gene editing offers superior specificity compared to older methods.
Key elements for successful genome engineering include inducing DNA breaks and accurate detection of modifications.

Conclusions:

Genetic engineering principles are evolving with technological advancements.
CRISPR/Cas9 has transformed genome engineering, largely replacing previous endonuclease tools.
Future research aims to enhance CRISPR/Cas9 efficiency, particularly homology-directed repair.