Lipid nanoparticle-mediated mRNA delivery system into preimplantation embryos†

Area of Science:

• Developmental Biology
• Gene Delivery
• Nanotechnology

Background:

• Lipid nanoparticles (LNPs) are advanced nonviral vectors for mRNA delivery.
• Efficiently delivering mRNA into early-stage embryos is crucial for studying developmental processes.

Purpose of the Study:

• To evaluate lipid nanoparticles (LNPs) as a delivery system for mRNA into preimplantation embryos.
• To assess the efficiency and toxicity of LNPs for gene transfer in early embryonic development.

Main Methods:

• Synthesized and characterized LNPs with varying cholesterol content for surface polarity.
• Delivered enhanced green fluorescent protein (EGFP) and Cre recombinase mRNA via LNPs into mouse zygotes and blastocysts.
• Assessed mRNA delivery efficiency, gene expression (EGFP), and recombination (mTmG reporter) at different embryonic stages (blastocyst, E12.5).

Main Results:

• Neutral or anionic cholesterol-based LNPs showed lower toxicity and better suitability for zygote transduction.
• LNP-mediated mRNA delivery resulted in high percentages of EGFP expression in embryos at blastocyst and E12.5 stages.
• Blastocyst stage embryo treatment led to trophectoderm-specific EGFP expression, with subsequent placenta-specific expression observed at E12.5.

Conclusions:

• Lipid nanoparticles represent a novel and effective method for mRNA delivery into preimplantation embryos.
• This LNP-based approach facilitates gene function studies across pre-, peri-, and post-implantation embryonic development.
• LNPs show promise for advancing research in reproductive biology and developmental toxicology.