A method for gene knockdown in the retina using a lipid-based carrier.

Area of Science:

• Ophthalmology
• Molecular Biology
• Gene Therapy

Background:

• Small non-coding nucleic acids like siRNA and miRNA are crucial for understanding gene function and developing therapies for neurodegenerative diseases.
• Effective delivery and transfection of nucleic acids into the retina are essential for these applications.

Purpose of the Study:

• To investigate Invivofectamine 3.0, a lipid-based transfection agent, for its efficacy in delivering nucleic acids to the retina.
• To assess the safety and transfection efficiency of Invivofectamine 3.0 in rodent models.

Main Methods:

• Intravitreal injections of Invivofectamine 3.0 with various siRNAs (scrambled, Gapdh, Il-1β, C3) or PBS in rodents.
• Histological analysis (TUNEL, IBA1), qPCR for gene expression, electroretinography (ERG), and optical coherence tomography (OCT) to evaluate retinal health and transfection.
• Visualization of transfection using fluorescently labeled siRNA.

Main Results:

• Invivofectamine 3.0 alone did not induce significant retinal cell death, inflammation, or changes in gene expression, ERG, or OCT.
• Targeted gene knockdown of Gapdh, Il-1β, and C3 was achieved in the retina for up to 4 days post-injection.
• Fluorescent siRNA demonstrated transfection throughout the neural retina, including ganglion, inner nuclear, and outer nuclear layers.

Conclusions:

• Invivofectamine 3.0 is a safe and effective transfection agent for delivering nucleic acids to the retina.
• This method supports gene function studies and holds potential for developing retinal therapeutics.