This article describes a method for delivering plasmid DNA encoding interleukin-15 into the skin of genetically modified mice using acupuncture techniques. The procedure aims to facilitate DNA uptake by skin cells, including keratinocytes and dendritic cells, through a process called acufection.
Prepare an anesthetized genetically modified mouse lacking cytokine interleukin-15 expression.
Shave the dorsal flank skin and apply a hair removal cream to eliminate any remaining hair shafts, facilitating improved DNA infusion into the skin in subsequent steps.
Sterilize the dorsal skin surface and mark the target area for DNA transfer.
Apply a small drop of plasmid DNA encoding interleukin-15 to the marked surface.
Using an acupuncture needle bundle, repeatedly prick the marked skin surface.
The pricking motion of the needle bundle gently oscillates the skin surface, loosening the dead cells of the outermost skin layer.
Further, the oscillating needles scrape the plasma membranes of cells, promoting acufection, or acupuncture-mediated DNA delivery into the cells, including keratinocytes and dendritic cells.
Now, allow the mouse to recover.
Successfully transfected cells in the skin produce interleukin-15 proteins.
After anesthetizing the mouse, apply veterinary ophthalmological ointment to its eyes to prevent dryness. Then, shave the skin on the back of the animal and apply a depilatory cream to dissolve keratin proteins in the remaining hair shafts. Following depilation, wipe off the cream with water-soaked cotton.
Using a sterile cotton swab soaked in 70% ethanol, disinfect the skin's surface and mark the target skin area with a previously prepared template. Apply 10 microliters of the DNA solution containing 10 micrograms of plasmid DNA to the desired skin area. To acufect the mouse, hold the acupuncture apparatus perpendicularly to the skin, and repetitively puncture the designated surface either 100 times over 30 seconds or until the solution applied to the skin absorbs.
To achieve successful plasmid DNA expression in the skin tissue, it is important to avoid cutting deep into the skin and to make sure the DNA solution is well-absorbed.
After completing the procedure, place the acufected mouse on a heating pad to maintain its body temperature until recovery from anesthesia. When the mouse regains sufficient consciousness, return it to its cage and provide water and food. Closely monitor the mouse for any symptoms of discomfort, including behavioral changes, abnormal appearance for the first 48 hours after the procedure.
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