Targeted genomic translocations and inversions generated using a paired prime editing strategy

Area of Science:

• Genetics
• Molecular Biology
• Cancer Research

Background:

• Chromosomal rearrangements are common in cancers, often creating fusion oncogenes.
• Engineered nucleases like CRISPR-Cas9 can model these rearrangements but cause unwanted mutations.

Purpose of the Study:

• To develop a novel method for precise chromosomal translocation and inversion.
• To overcome limitations of existing gene editing tools for disease modeling.

Main Methods:

• Developed prime editor nuclease-mediated translocation and inversion (PETI) using prime editor 2 (PE2) nuclease and paired pegRNAs.
• Applied PETI to episomal reporters and human cells to induce DNA recombination and chromosomal translocations.
• Created cancer-associated translocations (NPM1-ALK, EML4-ALK) using PETI.

Main Results:

• PETI successfully introduced precise chromosomal translocations and inversions in human cells.
• The method demonstrated programmable control over DNA recombination.
• PETI achieved efficiencies comparable to Cas9 for generating chromosomal rearrangements.

Conclusions:

• PETI is a novel, programmable tool for generating chromosomal translocations and inversions.
• This method accurately models cancer-associated gene fusions.
• PETI holds potential for creating advanced disease models and applications in gene therapy.