Targeting bacteriophage to mammalian cell surface receptors for gene delivery
Targeting bacteriophage to mammalian cell surface receptors for gene delivery
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DOI：
10.1089/hum.1998.9.16-2393
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发表时间：
1998-11-01
影响因子：
4.2
通讯作者：
Baird, A
中科院分区：
文献类型：
--
作者：
Larocca, D;Witte, A;Baird, A
丝状噬菌体代表了自然界最优雅的包装和传递 DNA 的方式之一。为了开发通过噬菌体基因传递发现配体的新方法，我们通过将碱性成纤维细胞生长因子 (FCF2)、转铁蛋白或表皮生长因子 (EGF) 附加到丝状噬菌体的外壳蛋白并测量靶细胞中 CMV 启动子驱动的报告基因表达，赋予丝状噬菌体哺乳动物细胞向性。在该系统中，FGF2 是更有效的靶向细胞中绿色荧光蛋白 (GFP) 或 β-半乳糖苷酶 (β-Gal) 活性的检测需要 FGF2 靶向，并且依赖于噬菌体浓度。通过将靶向噬菌体与 FGF2 竞争、FGF2 靶向噬菌体无法转导高亲和力 FGF 受体阴性细胞以及它们转导这些相同细胞的能力，证明了靶向高亲和力 FGF 受体的特异性当用高亲和力 FGF 受体 FGFR1 稳定转染时，通过选择 G418 抗性集落建立了长期转基因表达，这表明，通过适当的靶向向性，丝状噬菌体可以作为将靶向基因递送至哺乳动物细胞的载体。
Filamentous bacteriophages represent one of nature's most elegant ways of packaging and delivering DNA, In an effort to develop novel methods for ligand discovery via phage gene delivery, we conferred mammalian cell tropism to filamentous bacteriophages by attaching basic fibroblast growth factor (FCF2), transferrin, or epidermal growth factor (EGF) to their coat proteins and measuring CMV promoter-driven reporter gene expression in target cells, In this system, FGF2 was a more effective targeting agent than transferrin or EGF, The detection of green fluorescent protein (GFP) or beta-galactosidase (beta-Gal) activity in cells required FGF2 targeting and was phage concentration dependent, Specificity of the targeting for high-affinity FGF receptors was demonstrated by competing the targeted phage with FGF2, by the failure of FGF2-targeted bacteriophage to transduce high-affinity FGF receptor-negative cells, and by their ability to transduce these same cells when stably transfected with FGFR1, a high-affinity FGF receptor, Long-term transgene expression was established by selecting colonies for G418 resistance, suggesting that with the appropriate targeted tropism, filamentous bacteriophage can serve as a vehicle for targeted gene delivery to mammalian cells.