Acufection-Mediated Delivery of Immune-Related Genes into Mouse Skin Tissues

Abstract

Source: Lin, Y., et al., Development of an Economical DNA Delivery System by "Acufection" and its Application to Skin Research. J. Vis. Exp. (2017)

The video demonstrates a method for directly transferring the interleukin-15 encoding plasmid DNA into the skin tissue of an interleukin-15 deficient mouse model through acupuncture using microneedles. These microneedles generate micropunctures in the outermost skin layer, facilitating the absorption and transfection of plasmid DNA into the epidermal cells, including keratinocytes, within the skin tissue.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Procedure for Acufection

NOTE: The optimal amount of DNA and the area of the target skin surface may vary for different genes and need to be further optimized. The pricking force and number of times to loosen the horny layer of the skin also may vary depending on the skin thickness. These conditions must be carefully determined after evaluating the expression level of acufected gene transcripts by qRT-PCR.

1. Weigh the mice a....

Results

Figure 1. Illustration of Acupuncture Needles. (A) The acupuncture needle (36 G x 0.5") is composed of a handle and a needle end. (B) Ten acupuncture needles are bound in a bundle using adhesive tapes. The length of the bundle is approximately 4 c.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Accu Handy Needle	Accu	36Gx0.5	Medical device
NairTM Lotion	Church & Dwight	N/A	Use to remove hair
Shandon Xyline substitute	Thermo Fisher Scientific	6764506	Deparaffinization
Avertin (2,2,2-Tribromethanol)	Sigma-Aldrich	T4,840-2	Anesthesia
2-methyl-2-butanol	Sigma-Aldrich	152463	Solvent for the dissolution of avertin
QIAGEN® Plasmid Midi Kit	QIAGEN	12143	Purification of plasmid DNA from E. coli
Mouse IL-15/IL-15R Complex ELISA Ready-SET-Go kit	eBioscience	88-7215	Measure IL-15 protein