Song Lin, Chau Lillian, Sakamoto Yoshitaka, Nakashima Juichiro, Koide Masafumi, Tuan Rocky S
Cartilage Biology and Orthopaedics Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892-8022, USA.
Mol Ther. 2004 Apr;9(4):607-16. doi: 10.1016/j.ymthe.2004.01.017.
Gene delivery is an essential research tool for elucidating gene structure, regulation, and function in biomedical research and is the technological basis for gene therapy. However, the application of nonviral vectors in mammalian cell transfection and gene therapy is limited in that current methods require large amounts of exogenous DNA and/or exhibit high cytotoxicity and low transfection efficiency in primary cells. Here we describe the development of a novel, noninvasive gene delivery protocol using plasmid DNA vectors, based on the principle of electric field-induced molecular vibration. This method enables foreign DNA molecules to penetrate the plasma membrane and enter the cytoplasm of both primary mesenchymal progenitor cells and established cell lines of various species, at high efficiency and with low cell mortality. This procedure requires no special reagents, allows stable expression of transduced DNA, and does not interfere with the normal cellular differentiation activities of human and chick mesenchymal progenitors.
基因递送是生物医学研究中阐明基因结构、调控和功能的重要研究工具，也是基因治疗的技术基础。然而，非病毒载体在哺乳动物细胞转染和基因治疗中的应用受到限制，因为目前的方法需要大量的外源DNA，并且/或者在原代细胞中表现出高细胞毒性和低转染效率。在此，我们基于电场诱导分子振动的原理，描述了一种使用质粒DNA载体的新型非侵入性基因递送方案的开发。该方法能够使外源DNA分子高效穿透质膜并进入原代间充质祖细胞和各种物种的已建立细胞系的细胞质，同时细胞死亡率低。该程序不需要特殊试剂，允许转导的DNA稳定表达，并且不干扰人和鸡间充质祖细胞的正常细胞分化活动。
