https://scholars.lib.ntu.edu.tw/handle/123456789/414238
標題: | Development of an Economical DNA Delivery System by "Acufection" and its Application to Skin Research | 作者: | Lin, Yu-Jei Lee, Tsung-Lin CHIA-CHI KU |
關鍵字: | Immunology; Issue 122; animals; inflammation; skin; keratinocytes; DNA transfer; protein isoform; interleukin-15; imiquimod | 公開日期: | 2017 | 出版社: | JOURNAL OF VISUALIZED EXPERIMENTS | 卷: | 2017 | 期: | 122 | 來源出版物: | Journal of visualized experiments : JoVE | 摘要: | Dysregulation of immune response in skin is associated with numerous human skin disorders. Direct transfer of immune-related genes into skin tissue is a fascinating approach to investigate immune modulation of cutaneous inflammation in mouse models of human diseases. Here we present a cost-effective protocol that delivered naked DNA in mouse skin and leads to transgene expression. The method is coined "acufection", denoting acupuncture-mediated DNA transfection. To perform acufection, mouse skin was first infused with DNA in phosphate-buffered saline (PBS) and then pricked lightly with a bundle of acupuncture needles to facilitate the absorption of DNA and transfection into cells. The plasmid DNA is presumably taken up by the keratinocyte and dendritic cells (DCs) in the skin and expressed into protein. Mechanical prick with the needles per se did not cause skin damage or induce keratinocyte activation. The expression of the transfected genes was detected in the skin at both transcriptional and translational levels following acufection for 2 days and maintained up to 7 days. The primary goal for the development of this acufection method was to investigate a previously undefined isoform of IL-15. Using this method, an alternatively spliced IL-15 isoform with partially deleted exon 7 (IL-15ΔE7) was expressed in the skin and subsequently treated with a Toll-like receptor 7 (TLR7) agonist, imiquimod (IMQ), to induce inflammation. Acufection-delivered IL-15ΔE7 in skin suppressed keratinocyte proliferation, epidermal thickness and neutrophil recruitment in IMQ-induced cutaneous inflammation. With increasing interest in identifying the regulatory mechanisms of cutaneous inflammation, the protocol described here provides a cost effective and versatile alternative to the gene gun system or microseeding for DNA delivery in vivo. It may potentially allow discovery of the function of a novel gene in the skin or for investigating new treatment for cutaneous diseases. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85017633985&doi=10.3791%2f55206&partnerID=40&md5=0c2e0c031eae2c06581bd070540fa91c https://scholars.lib.ntu.edu.tw/handle/123456789/414238 |
ISSN: | 1940-087X | DOI: | 10.3791/55206 |
顯示於: | 免疫學研究所 |
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