Cation substitution in cationic phosphonolipids:: A new concept to improve transfection activity and decrease cellular toxicity
Cation substitution in cationic phosphonolipids:: A new concept to improve transfection activity and decrease cellular toxicity
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DOI：
10.1021/jm000006z
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发表时间：
2000-11-30
影响因子：
7.3
通讯作者：
Férec, C
中科院分区：
文献类型：
--
作者：
Floch, V;Loisel, S;Férec, C
阳离子脂质已被证明是一种有趣的替代病毒载体介导的基因传递到体外和体内模型应用。先前的研究表明，即使脂质疏水结构域或脂质极性结构域的微小结构修饰也会导致基因传递效率的显着变化。在此之前，我们开发了一类新的阳离子脂质，称为阳离子磷脂，并描述了这些载体将DNA转移到不同细胞系和体内的能力。到目前为止，在所有新的阳离子脂质中，氮原子总是携带阳离子或多阳离子电荷。最近我们开发了一系列新的阳离子磷酸脂，其特征是磷或砷原子带正电荷。在第二步中，我们还研究了阳离子和疏水结构域之间的连接体长度对转染活性的影响。利用荧光素酶报告基因，研究了该新型阳离子磷脂文库在不同细胞系（HeLa、CFT1、K562）和体内的转染活性。荧光法检测荧光素酶的表达。我们证明了阳离子磷脂（N -> P或As）极性域的阳离子取代导致体外和体内转染活性的显著增加和细胞毒性的降低。
Cationic lipids have been shown to be an-interesting alternative to viral vector-mediated gene delivery into in vitro and in vivo model applications. Prior studies have demonstrated that even minor structural modifications of the lipid hydrophobic domain or of the lipid polar domain result in significant changes in gene delivery efficiency. Previously, we developed a novel class of cationic lipids called cationic phosphonolipids and described the ability of these vectors to transfer DNA into different cell lines and in vivo. Up until now, in all new cationic lipids, nitrogen atoms have always carried the cationic or polycationic charge. Recently we have developed a new series of cationic phosphonolipids characterized by a cationic charge carried by a phosphorus or arsenic atom. In a second step, we have also examined the effects of the linker length between the cation and the hydrophobic domain as regards transfection activity. Transfection activities of this library of new cationic phosphonolipids were studied in vitro in different cell lines (HeLa, CFT1, K562) and in vivo using a luciferase reporter gene. A luminescent assay was carried out to assess luciferase expression. We demonstrated that cation substitution on the polar domain of cationic phosphonolipids (N --> P or As) results in significant increase in transfection activity for both in vitro and in vivo assays and decrease of cellular toxicity.