Electroporation-Mediated Transfection of Mouse Dorsal Root Ganglia to Promote Sensory Axon Regeneration

Take an anesthetized mouse with exposed L4 and L5 dorsal root ganglia containing sensory neuron cell bodies.

Load a microinjection pipette with fluorophore-tagged RNA constructs that regulate axon regeneration and a tracer dye.

Inject the mixture into the DRGs.

Using electroporation electrodes, apply electric pulses to the DRGs.

This disrupts the membranes of sensory neuron cell bodies, creating pores that allow the constructs to enter the cytoplasm, facilitating transfection.

Suture the incision and allow the mouse to recover.

After a few days, re-anesthetize and secure the mouse.

Make an incision lateral to the midline on the left. Dissect the muscles to expose the sciatic nerve.

Crush the nerve to injure sensory neuron axons. Mark the site with a suture knot.

Suture the incision and allow the mouse to recover.

The transfected constructs target mRNAs in DRG sensory neuron cell bodies, transiently modulating protein production to enhance sensory axon regeneration post-injury.

For DRG injection, load the DNA plasmids or RNA oligos into the glass capillary pipette. Then insert the tip of the capillary glass pipette carefully into the DRG. And gradually inject 1 microliter solution of DNA plas