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In vivo electroporation-based chromosomal engineering in murine uterine epithelium

Iwata, S.; Miura, Y.; Iwamoto, T.

2025-03-01 genomics
10.1101/2025.02.25.640089 bioRxiv
Show abstract

Clustered regularly interspaced palindromic repeat (CRISPR)/CRISPR-associated protein (Cas)-based in vivo chromosomal rearrangements are a promising approach for generating model organisms with specific chromosomal abnormalities. However, conventional in vivo methods rely on viral vectors, which are expensive, require specialized equipment, and pose potential safety risks, thereby limiting their widespread application. To overcome the limitations above, we developed a novel, efficient, and cost-effective in vivo chromosomal engineering strategy using CRISPR ribonucleoprotein electroporation for the murine uterine epithelium. Our method successfully induced translocations at multiple loci and repaired a 57.8-Mb inversion. The findings of the present study establish in vivo electroporation as a practical alternative to traditional chromosomal engineering methods and provide a foundation for its broader application in genome editing technologies.

Published in PLOS ONE · training set

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