Related Experiment Video
Updated: May 25, 2026
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
Oligonucleotide-directed gene-editing technology: mechanisms and future prospects
Ioannis Papaioannou1, J Paul Simons, James S Owen
1UCL Medical School, Division of Medicine (Upper 3rd Floor), Royal Free Campus, Rowland Hill Street, London NW3 2PF, UK.
Expert Opinion on Biological Therapy
|February 11, 2012
Summary
Single-stranded DNA oligonucleotides (ssODNs) enable precise gene editing in mammalian cells. This technology, combined with nucleases, is advancing therapeutic applications and in vivo gene editing possibilities.
Area of Science:
- Molecular Biology
- Genetics
- Genomics
Background:
- Gene editing utilizes synthetic oligonucleotides for site-specific genomic modifications.
- Single-stranded DNA oligonucleotides (ssODNs) have evolved from earlier RNA-DNA chimeraplasts into a reliable gene-editing tool with therapeutic potential.
Purpose of the Study:
- To review the mechanisms linking gene editing to DNA repair and replication systems.
- To highlight advancements in ssODN chemistry and their combination with endonuclease technologies for enhanced genome editing.
Main Methods:
- Exploration of the roles of homologous recombination (HR), nucleotide excision repair (NER), and mismatch repair (MMR) in gene editing.
- Investigation of ssODN chemistries and their impact on editing efficiency and cell viability.
- Examination of the synergistic use of ssODNs with zinc finger nucleases for high-frequency genome editing.
Main Results:
- While HR and NER are implicated, MMR can inhibit gene editing.
- Optimized ssODN chemistries improve editing efficiency and cell viability, mitigating cell cycle arrest.
- Tandem use of ssODNs with zinc finger nucleases significantly enhances genome editing frequency.
Conclusions:
- Gene editing via ssODNs has progressed from reporter systems to endogenous gene modification.
- Improved understanding of mechanisms, ssODN design, and endonuclease combinations are driving advancements.
- Routine in vitro and ex vivo success, including in embryonic stem (ES) and induced pluripotent stem (iPS) cells, points to future in vivo organ gene editing applications.
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