Subretinal Injection of Viral Vectors into the Retinal Pigment Epithelium in a Mouse Model

Begin with an anesthetized mouse and apply a pupil-dilating agent.

Under a microscope, open the eyelid and slightly protrude the eye to expose the injection site.

Apply a drop of ophthalmic viscoelastic solution, then place a coverslip to visualize the retina.

Using a fine needle, create a hole behind the cornea-sclera junction, avoiding damage to the blood vessels and the lens.

Through this pre-punctured hole, insert a blunt syringe tangentially into the subretinal space near the retinal pigment epithelium or RPE, which supports retinal neurons.

Inject the lentiviral vector carrying target genes into the subretinal space.

Observe the formation of a subretinal bleb due to fluid accumulation, confirming successful vector delivery.

Gently close the eyelid to cover the injection site and allow the mouse to recover.

The lentivirus enters the RPE via endocytosis, releases its genetic material, and converts it into the corresponding DNA, which integrates with the host genome, enabling gene expression.

First, prepare a microliter syringe with a 33-gauge blunt needle. Sterilize it using ethylene oxide gas. Next, prepare a dilution of the viral vectors in PBS in a micro fuge tube at about 1 million