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March 25, 2026Communications BiologyOpen Access

Optimizing CRISPR precision in mouse embryos via microhomology-mediated end joining-dominant targeting

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Authors

KLKhanui LkhagvadorjEOEiichi OkamuraTTTaito Taki

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Overview

This hybrid approach enhances gene editing predictability in mice, suggesting improved outcomes in genetic studies.

Key Points

  • To optimize CRISPR/Cas9 gene editing precision in mouse embryos using MMEJ-dominant targeting.
  • Developed a hybrid genome editing strategy combining in silico prediction and in vitro validation.
  • Applied the approach to mouse embryonic stem cells (mESCs) for testing guide RNAs (gRNAs).
  • Pre-selected gRNAs predicted to enhance microhomology-mediated end joining (MMEJ) for gene targeting.
  • Injected validated gRNAs into mouse embryos targeting Tyr and Fgf10 genes.
  • Efficient generation of F0 mice with uniform genotypes was achieved.
  • Mutation patterns in mESCs closely matched those in blastocysts and post-implantation embryos.
  • The strategy improved predictability and reproducibility of genome editing outcomes.

Cite This Study

Lkhagvadorj et al. (2026) studied this question.

synapsesocial.com/papers/69c37acab34aaaeb1a67cb49https://doi.org/10.1038/s42003-026-09771-z
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