A Highly Efficient siRNA Transfection Method in Primary Cultured Cortical Neurons

Area of Science:

• Neuroscience
• Molecular Biology
• Biotechnology

Background:

• Neuron transfection is challenging due to cell sensitivity and cytotoxicity of conventional methods like Lipofectamine.
• Lentiviral transfection is efficient but costly and complex.
• A novel nanoparticle-based lipid reagent for oligonucleotide delivery exists but its neuronal efficacy is unproven.

Purpose of the Study:

• To evaluate the efficacy of a novel nanoparticle-based lipid reagent for small interfering RNA (siRNA) transfection in primary cultured mouse cortical neurons.
• To establish a simple, efficient, and non-toxic method for in vitro neuronal transfection.

Main Methods:

• Utilized a novel siRNA-specific transfection reagent from Kermey, a nanoparticle-based lipid material.
• Transfected primary cultured mouse cortical neurons with Phosphoglycerate dehydrogenase (PHGDH) siRNA and negative control siRNA (siNC) after 72 hours of in vitro culture.
• Performed transfection without medium replacement 6-8 hours post-transfection.

Main Results:

• Achieved robust and efficient in vitro transfection of siRNA into primary cultured mouse cortical neurons.
• Observed no adverse effects in transfected neurons throughout the experimental period.
• Demonstrated consistent and efficient knockdown of the target protein.

Conclusions:

• The novel nanoparticle-based reagent provides an efficient and safe method for siRNA delivery into primary neurons.
• This streamlined protocol minimizes cellular stress and simplifies the transfection workflow.
• The method enables reliable gene silencing in neuronal cells for research applications.