Acufection-Mediated Delivery of Immune-Related Genes into Mouse Skin Tissues
Published: May 31, 2024
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 …
Representative 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 cm as measured by the ruler (lower panel).
Divulgations
The authors have nothing to disclose.
Materials
| 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 |
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Citer Cet Article
Acufection-Mediated Delivery of Immune-Related Genes into Mouse Skin Tissues. J. Vis. Exp. (Pending Publication), e22256, doi: (2024).