Wagner E, Zenke M, Cotten M, Beug H, Birnstiel M L
Research Institute of Molecular Pathology, Vienna, Austria.
Proc Natl Acad Sci U S A. 1990 May;87(9):3410-4. doi: 10.1073/pnas.87.9.3410.
We have developed a high-efficiency nucleic acid delivery system that uses receptor-mediated endocytosis to carry DNA macromolecules into cells. We accomplished this by conjugating the iron-transport protein transferrin to polycations that bind nucleic acids. Human transferrin, as well as the chicken homologue conalbumin, has been covalently linked to the small DNA-binding protein protamine or to polylysines of various sizes through a disulfide linkage. These modified transferrin molecules maintain their ability to bind their cognate receptor and to mediate efficient iron transport into the cell. The transferrin-polycation molecules form electrophoretically stable complexes with double-stranded DNA, single-stranded DNA, and modified RNA molecules independent of nucleic acid size (from short oligonucleotides to DNA of 21 kilobase pairs). When complexes of transferrin-polycation and a bacterial plasmid DNA containing the gene for Photinus pyralis luciferase are supplied to eukaryotic cells, high-level expression of the luciferase gene occurs, demonstrating transferrin receptor-mediated endocytosis and expression of the imported DNA. We refer to this delivery system as "transferrinfection."
我们开发了一种高效核酸递送系统，该系统利用受体介导的内吞作用将DNA大分子转运到细胞中。我们通过将铁转运蛋白转铁蛋白与结合核酸的聚阳离子偶联来实现这一点。人转铁蛋白以及鸡的同源物伴清蛋白已通过二硫键与小DNA结合蛋白鱼精蛋白或各种大小的聚赖氨酸共价连接。这些修饰的转铁蛋白分子保持其结合同源受体并介导铁高效转运到细胞中的能力。转铁蛋白 - 聚阳离子分子与双链DNA、单链DNA和修饰的RNA分子形成电泳稳定的复合物，而与核酸大小无关（从短寡核苷酸到21千碱基对的DNA）。当将转铁蛋白 - 聚阳离子与含有萤火虫荧光素酶基因的细菌质粒DNA的复合物提供给真核细胞时，荧光素酶基因会高水平表达，这证明了转铁蛋白受体介导的内吞作用以及导入DNA的表达。我们将这种递送系统称为“转铁感染”。
