Peer-reviewed veterinary case report
How to study gene effects on mouse limb development with siRNA
By Yea JH et al.·2026·Department of Biomedical Engineering, United States·View original on Europe PMC →
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Original publication title: Protocol to investigate the effects of localized gene expression disruption on mouse limb development via direct siRNA injection and explant culture.
Plain-English summary
This study outlines a method for reducing the activity of specific genes in the hind limbs of developing mouse embryos. Researchers inject a special substance called siRNA into the limb tissue to see how it affects growth and development. They also describe how to prepare and maintain these tissue samples for observation and analysis. This technique allows scientists to carefully control gene activity and study its effects on limb and joint formation. The approach is aimed at better understanding how limbs develop in mice.
Abstract
Here, we present a protocol for localized gene knockdown in embryonic mouse hindlimb explants using direct small interfering RNA (siRNA) injection, combined with a semi-quantitative morphology scoring system to assess developmental outcomes. We describe steps for mating of samples, preparing hindlimb explants for culture and siRNA-Lipofectamine complex, and direct injection of the siRNA-Lipofectamine complex. We then detail procedures for explant culture and maintenance, data analysis, tissue fixation, and cryoprotection. This protocol approach enables spatially precise modulation of gene expression during limb and joint development.
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Search related cases →Original publication on Europe PMC: https://europepmc.org/article/MED/41764661