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Related Experiment Video

Updated: Jun 20, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

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Efficient Genome Editing with Chimeric Oligonucleotide-Directed Editing.

Long T Nguyen^1,2,3, Noah R Rakestraw⁴, Brianna L M Pizzano⁵

¹Department of Molecular Biology, Princeton University, Princeton, NJ, USA.

Biorxiv : the Preprint Server for Biology

|July 19, 2024

Summary

This summary is machine-generated.

New chimeric oligonucleotide-directed editing (CODE) systems offer precise, search-and-replace genome editing. These novel editors achieve efficient gene modification in human cells with minimal unintended edits, expanding genome editing capabilities.

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Prime editing represents a significant advancement in precise genome editing.
Existing Cas9-based methods have limitations that newer approaches aim to overcome.

Purpose of the Study:

To develop novel genome editing systems for precise, search-and-replace gene modifications.
To engineer fusion proteins that enhance editing efficiency and specificity.

Main Methods:

Development of chimeric oligonucleotide-directed editing (CODE) systems using nCas9-DNA polymerase fusion proteins.
Engineering of CODEMax and CODEMax(exo+) editors incorporating a Bst DNA polymerase derivative.
Evaluation of editing efficiency and unintended edits in human cells.

Main Results:

CODEMax and CODEMax(exo+) demonstrated efficient genome modifications in human cells.
These CODE systems exhibited low levels of unintended edits.
CODEs showed improved efficiency for small insertions, deletions, and substitutions compared to PEMax at various loci.

Conclusions:

CODE systems provide a powerful new tool for genome manipulation.
These editors complement existing prime editing technologies.
CODEs enable precise editing without inducing double-stranded breaks, expanding the genome editing toolbox.