Efficient transfer of genetic material into mammalian cells using Starburst polyamidoamine dendrimers
Efficient transfer of genetic material into mammalian cells using Starburst polyamidoamine dendrimers
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DOI：
10.1073/pnas.93.10.4897
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发表时间：
1996-05-14
影响因子：
11.1
通讯作者：
Baker, JR
中科院分区：
文献类型：
--
作者：
KukowskaLatallo, JF;Bielinska, AU;Baker, JR
星型聚酰胺-胺树枝状大分子是一类新型的合成高分子，具有独特的结构和物理性质。研究了这些聚合物在多种哺乳动物细胞系中结合DNA和增强DNA转移和表达的能力。二十种不同类型的聚酰胺-胺树枝状聚合物的合成，并使用定义明确的分析技术确认的聚合物结构。使用两种报告基因系统：萤火虫荧光素酶和细菌β-半乳糖苷酶，使用树枝状聚合物的质粒DNA转染的效率进行了检查。使用各种树枝状聚合物进行转染，并测定报告蛋白的表达水平。使用DNA/树枝状聚合物复合物以最小的细胞毒性实现了广泛的真核细胞和细胞系的高效转染。然而，抑制细胞的能力仅限于某些类型的树枝状聚合物，并且在某些情况下需要存在额外的化合物，如DEAE-葡聚糖，这似乎会改变复合物的性质。一些细胞系表现出增强的转染与氯喹的加入，表明内体定位的复合物。树枝状聚合物转染细胞的能力似乎取决于聚合物表面上伯氨基的大小、形状和数量。然而，在实现转染方面最有效的特定树枝状聚合物在不同类型的细胞之间变化。这些研究表明，Starburst树枝状聚合物可以在体外转染多种细胞类型，并提供了一种有效的方法来产生永久转染的细胞系。
Starburst polyamidoamine dendrimers are a new class of synthetic polymers with unique structural and physical characteristics. These polymers were investigated for the ability to bind DNA and enhance DNA transfer and expression in a variety of mammalian cell lines. Twenty different types of polyamidoamine dendrimers were synthesized, and the polymer structure was confirmed using well-defined analytical techniques. The efficiency of plasmid DNA transfection using dendrimers was examined using two reporter gene systems: firefly luciferase and bacterial beta-galactosidase. The transfections were performed using various dendrimers, and levels of expression of the reporter protein were determined. Highly efficient transfection of a broad range of eukaryotic cells and cell lines was achieved with minimal cytotoxicity using the DNA/dendrimer complexes. However, the ability to transfect cells was restricted to certain types of dendrimers and in some situations required the presence of additional compounds, such as DEAE-dextran, that appeared to alter the nature of the complex. A few cell lines demonstrated enhanced transfection with the addition of chloroquine, indicating endosomal localization of the complexes. The capability of a dendrimer to transfect cells appeared to depend on the size, shape, and number of primary amino groups on the surface of the polymer, However, the specific dendrimer most efficient in achieving transfection varied between different types of cells. These studies demonstrate that Starburst dendrimers can transfect a wide variety of cell types in vitro and offer an efficient method for producing permanently transfected cell lines.