Agarose-Embedded Electroporation-Assisted Gene Transfer in Mouse Cortical Interneuron Progenitors

Fill a micropipette with a DNA mixture containing plasmid vectors carrying the gene of interest and a tracking dye. Secure it in a micromanipulator.

Position an agarose block under the microscope and place an agarose-embedded mouse embryonic brain slice with cortical interneurons over it.

Lower the micropipette and inject the DNA mixture into the interneuron progenitor region. The tracking dye marks the injection site.

Position another agarose block on a Petri dish electrode and an agarose column on the cover electrode.

Transfer the slice onto the block and align the cover electrode over the injection site.

Apply brief electrical pulses to permeabilize cell membranes temporarily, allowing the plasmids to enter the nucleus.

Incubate the slice in a medium to maintain cell viability.

Inside the nucleus, the plasmid triggers gene expression, produces the target protein, and modulates interneuron progenitor functions.

For the injection, mix the expression and control vector DNA at a 1 microgram per microliter concentration for each vector, and add fast green stock solution at a 1 to 10th dilution. Fill a pulled 0.5 millimeter inner diameter, 1 millimeter outer diameter glass micropipette with