Poly(amidoamine)s as potential nonviral vectors: Ability to form interpolyelectrolyte complexes and to mediate transfection in vitro
Poly(amidoamine)s as potential nonviral vectors: Ability to form interpolyelectrolyte complexes and to mediate transfection in vitro
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DOI：
10.1021/bm010079f
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发表时间：
2001-09-01
影响因子：
6.2
通讯作者：
Duncan, R
中科院分区：
文献类型：
--
作者：
Richardson, SCW;Pattrick, NG;Duncan, R
聚（酰胺基胺）（PAA）是水溶性聚合物，显示pH依赖性膜活性。PAA有可能作为融合肽的合成替代物，从而促进内体逃逸。本研究的目的是首次研究PAA是否具有复合DNA、保护DNA免受核酸酶降解以及促进体外转染的能力。研究了PAAs伊萨I（M-n 6900）和伊萨23（M-n 10500）及其2-苯乙胺类似物伊萨4和伊萨22（M-n近似于8000）。所有的PAA都延缓λ Hind III DNA的电泳迁移率，证明了插入复合物（IPEC）的形成，并且使用TEM可见直径为80-150 nm的环状体（10：1聚合物过量）。观察到DNase 11抑制。在聚合物：DNA比率为10：1时，这是伊萨1（89.6 +/- 6.1%）、伊萨4（92.2 +/- 11.2%）、伊萨22（69.4 +/- 3.7%）和伊萨23（58.0 +/- 10.0%）。PAAs显示出介导pSV β-半乳糖苷酶转染HepG 2细胞的能力。在载体与DNA质量比为5：1时，伊萨23显示出与聚乙烯亚胺和LipofectIN相当的转染能力，并且比LipofectACE更有效。这些性质表明，PAA值得进一步开发为内体溶解载体。
Poly(amidoamine)s (PAAs) are water-soluble polymers that display pH-dependent membrane activity. PAAs have the potential to act as a synthetic alternative to fusogenic peptides and thus promote endosomal escape. The purpose of this study was to investigate for the first time whether PAA have the ability to complex DNA, protect it from nuclease degradation and to promote transfection in vitro. PAAs ISA I (M-n 6900) and ISA 23 (M-n 10 500) and their 2-phenylethylamine containing analogues ISA 4 and ISA 22 (M-n similar to8000) were studied. All PAAs retarded the electrophoretic mobility of lambda Hind III DNA demonstrating interpolyelectrolyte complex (IPEC) formation and toroids of 80-150 nm in diameter (10: 1 polymer excess) were visible using TEM. DNase 11 inhibition was observed. At a polymer:DNA ratio of 10: 1, this was ISA 1(89.6 +/- 6.1%), ISA 4 (92.2 +/- 11.2%), ISA 22 (69.4 +/- 3.7%), and ISA 23 (58.0 +/- 10.0%). PAAs demonstrated the ability to mediate pSV beta-galactosidase transfection of HepG2 cells. At a vector:DNA mass ratio of 5:1, ISA 23 showed equivalent transfection ability compared with polyethylenimine and LipofectIN and was more effective than LipofectACE. These properties suggest that PAAs warrant further development as endosomolytic vectors.