Efficient genome editing with chimeric oligonucleotide-directed editing
Year: 2026
Type: article
Abstract: Prime editing has emerged as a precise and powerful genome editing tool, offering a favorable gene editing profile compared to other Cas9-based approaches. Here we report new nCas9-DNA polymerase fusion proteins to create chimeric oligonucleotide-directed editing (CODE) systems for search-and-replace genome editing. Through successive rounds of engineering, we developed CODEMax and CODEMax(exo+) editors that achieve efficient genome modifications in human cells with low unintended edits. CODEMax and CODEMax(exo+) contain an engineered Bst DNA polymerase derivative known for its robust strand displacement ability. Additionally, CODEMax(exo+) features a 5’ to 3’ exonuclease activity that promotes effective strand invasion and repair outcomes favoring the incorporation of the desired edit. We demonstrate CODEs can perform small insertions, deletions, and substitutions with improved efficiency compared to PEMax at many loci. Overall, CODEs complement existing prime editors to expand the toolbox for genome manipulations without double-stranded breaks.
Source: Nature Communications
Authors: Long T. Nguyen, Noah R. Rakestraw, Brianna L. M. Pizzano, Rajan Iyyappan, Cullen B. Young, Yujia Huang, Kate T. Beerensson, Anne Fang, Sydney G. Antal, Katerina V. Anamisis, Coleen M. D. Peggs, Jun Yan, Yangwode Jing, Jordan G. Lewis, Rebecca D. Burdine, Britt Adamson, Zongliang Jiang, Jared E. Toettcher, Cameron Myhrvold, Piyush K. Jain
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,990
Funders: CHDI Foundation, Princeton University, University of Florida, Searle Scholars Program, China Scholarship Council, National Institutes of Health, Centers for Disease Control and Prevention, University of Florida Health, International Centre for Bamboo and Rattan, National Human Genome Research Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: DNA Methylation at N6-Adenine in Placental Trophoblast Development, G4733222645, Chemistry-Biology Interface Training Program at the University of Florida, Control of the 4D chromatin landscape underlying gene activity during development, Methods and Logic in Molecular Biology Training Program, Discovery and engineering of CRISPR/Cas systems, A rapid CRISPR-based self-testing platform for early detection of HIV, Center for Genomic Editing and Recording: Development and Application of Next-Generation Genome and Epigenome Editing Methods to Advance the Study and Treatment of Human Disease, Causes and Consequences of DNA Replication Stress in the Mammalian Preimplantation Embryo
OpenAlex ID: W4400484366