Key Findings
The study demonstrates that lipid nanoparticle (LNP)-mediated delivery enables high-efficiency gene editing and phenotype generation in avian embryos with high viability. A primary result of this approach is the successful somatic genome editing of Runt-related transcription factor 2 (RUNX2) in chicken (Gallus gallus) embryos. This edit caused a severe loss and disruption of craniofacial ossification on the targeted side of the craniofacial skeleton, proving the platform’s ability to produce rapid functional in vivo gene interrogation.
Technical Details
The researchers developed a robust pipeline by optimizing LNP formulation, injection timing, and frequency. The platform supports the delivery of both plasmid DNA and RNA cargos. Specifically, the system can support the piggyBac transposon system, which allows for long-term labeling of cranial neural crest cells (NCCs) and persistent reporter expression. To achieve genome editing, the team utilized RNA-based CRISPR/Cas9 components targeting migrating cranial NCCs, which are critical contributors to the craniofacial skeleton.
Background and Context
Avian embryos are valuable models for developmental biology due to the ease of observing and manipulating fertilized eggs prior to hatching. However, traditional methods such as in ovo electroporation and viral transduction are limited by variable efficiency, high embryo mortality, and procedure-induced defects. These traditional vectors have often proven ineffective for the delivery of gene editing reagents like CRISPR/Cas9.
Significance and Outlook
Beyond the chicken embryo, the study confirms that LNPs can mediate transfection in domestic pigeon (Columba livia) and Pekin duck (Anas platyrhynchos) embryos. This versatility indicates that the LNP platform can be deployed across various avian species. The ability to achieve high-viability, reproducible gene manipulation positions this technology as a viable vector delivery platform for the study of avian development, disease, and evolution.
Source: https://www.biorxiv.org/content/10.64898/2026.10.13.753869v1
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