Hyndman Laura, Lemoine Jerome L, Huang Leaf, Porteous David J, Boyd A Christopher, Nan Xinsheng
Medical Sciences (Medical Genetics), Molecular Medicine Centre, The University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK.
J Control Release. 2004 Oct 19;99(3):435-44. doi: 10.1016/j.jconrel.2004.07.023.
The protein transduction domain (PTD) of the HIV-1 Tat protein can facilitate the cellular and nuclear uptake of macromolecular particles. Here, we demonstrate that incorporation without covalent linkage of a 17-amino acid PTD peptide into gene delivery lipoplexes improves gene transfer. Tat/Liposome/DNA (TLD) transfection, as evaluated by Fluorescence Activated Cell Scan analysis of a Green Fluorescence Protein expression plasmid, enabled transfection of highly recalcitrant primary cells in the form of air/liquid interface cultures of sheep tracheal epithelium. Treatment with chloroquine increased, and incubation at low temperature decreased, TLD transfection, suggesting that the endocytosis uptake pathway is involved.
HIV-1反式激活因子（Tat）蛋白的蛋白转导结构域（PTD）可促进大分子颗粒的细胞摄取和核摄取。在此，我们证明，将一种17个氨基酸的PTD肽非共价连接并入基因传递脂质体可改善基因转移。通过对绿色荧光蛋白表达质粒进行荧光激活细胞扫描分析评估的Tat/脂质体/DNA（TLD）转染，能够以羊气管上皮的气/液界面培养形式转染高度难转染的原代细胞。用氯喹处理可增强TLD转染，而在低温下孵育则降低TLD转染，这表明内吞摄取途径参与其中。
