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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

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Published on: March 12, 2018

Fourth-generation gene editors: Integration-based genome engineering.

Aidan E Kincaid¹, Alex W Hewitt^1,2,3,4, Rajendra Kc^1,2

¹Alphinia Bio, Lenah Valley, Hobart, TAS 7008, Australia.

Molecular Therapy. Advances

|May 27, 2026

Summary

This summary is machine-generated.

Fourth-generation gene editing tools, like integrase systems, offer precise, double-strand break-free DNA insertion. These advanced methods promise safer gene therapies for inherited diseases.

Keywords:

gene editors gene therapy genome engineering site-specific integration therapeutic gene integration

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

CRISPR-Cas9 gene editing, while revolutionary, can cause genomic instability due to double-strand breaks.
Innovation is needed to develop safer and more precise gene editing technologies.

Purpose of the Study:

To review advancements in fourth-generation gene editing technologies.
To focus on integrase systems enabling precise, double-strand break-free DNA insertion.

Main Methods:

Exploration of site-specific recombinases (tyrosine, large serine, bridge).
Introduction of DNA transposases (cut-and-paste systems, CRISPR-associated transposons).
Highlighting viral integrases and mobile group II introns/retrotransposons.

Main Results:

Diverse integrase systems facilitate efficient, site-specific, and DSB-free DNA insertion.
CRISPR-associated transposons offer RNA-guided, efficient, and directional insertion.
Viral integrases and mobile introns provide stable gene delivery and site-specific integration.

Conclusions:

Significant advances in efficiency, specificity, and delivery strategies have been achieved.
Refinement of integrase systems is crucial for next-generation gene-based therapies.
These tools hold potential for the definitive correction of inherited diseases.