Related Experiment Video
Updated: May 28, 2026
08:22
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
Published on: March 12, 2018
Fourth-generation gene editors: Integration-based genome engineering
Aidan E Kincaid1, Alex W Hewitt1,2,3,4, Rajendra Kc1,2
1Alphinia Bio, Lenah Valley, Hobart, TAS 7008, Australia.
Molecular Therapy. Advances
|May 27, 2026
Summary
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.
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.
Related Concept Videos
00:49What is Genetic Engineering?
Overview
01:28CRISPR/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...
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
02:31Homologous 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...