Expression of an epidermal growth factor-transdermal peptide fusion protein in Arabidopsis thaliana and its therapeutic effects on skin barrier repair

作者：Guangdong Yu, Shisheng Lin, Xulong Huang, Shuang Gao, Chengyang Song, Farid Khalilov, Qiongzhen Chen, Nipatha Issaro, Jiali Xiao, Xiashun Xu, Junchao Wang, Wengang Zhao, Yunpeng Wang, Nuo Xu · 发表于：Frontiers in Plant Science · 年份：2025 · DOI：10.3389/fpls.2025.1573193 · 被引用次数：4 · 研究领域：Transgenic Plants and Applications、Toxin Mechanisms and Immunotoxins、Plant Reproductive Biology

Epidermal growth factor (EGF) is recognized for its role in regulating keratinocyte proliferation and differentiation, thereby facilitating the restoration of impaired skin barriers. Nevertheless, challenges related to the penetration and safety of EGF remain to be resolved. In this study, we evaluated the efficacy of TDP1, a transdermal peptide, in enhancing the penetration of EGF through murine skin, utilizing EGF expressed in A. thaliana . The coding sequences of the TDP1 and EGF genes were cloned as a fusion construct into a plant expression vector. The resulting plasmid, pGM3301-TDP1-EGF, was introduced into A. thaliana via the floral dip method. Positive clones were identified using polymerase chain reaction (PCR). High-expression strains were selected through Western-blot analysis and enzyme-linked immunosorbent assay (ELISA). Homozygotes plants were obtained through self-pollination. The impact of the TDP1-EGF fusion protein on the restoration of a compromised epidermal barrier was assessed using dermatoscopy. Keratinocyte (KC) proliferation was examined via hematoxylin and eosin (H&E) staining, while KC differentiation, lipid synthesis, and inflammatory factors were analyzed using reverse transcription quantitative PCR (RT-qPCR) and immunohistochemistry. Compared to other expression systems, the A. thaliana system utilized for TDP1-EGF expression offers the advantages of being devoid of toxicity from endogenous plant substances, rendering it both safe and sui...