Lipoic acid-derived amphiphiles for redox-controlled DNA delivery
Lipoic acid-derived amphiphiles for redox-controlled DNA delivery
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DOI：
10.1016/s1074-5521(00)00030-2
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发表时间：
2000-10-01
影响因子：
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通讯作者：
Chroboczek, J
中科院分区：
文献类型：
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作者：
Balakirev, M;Schoehn, G;Chroboczek, J
背景：细胞内游离DNA从载体复合物中释放是限制非病毒基因递送效率的关键步骤之一。复合物应该足够稳定以防止DNA降解，但它应该在细胞内不稳定以允许DNA释放和转录。合成载体的去稳定化和降解也是降低其细胞毒性和增加转染细胞寿命所必需的。在这里，我们描述了由天然维生素原、硫辛酸制成的新的阳离子两亲物，其可逆地结合和释放DNA，这取决于硫辛酸部分的氧化还原状态。在氧化状态下，这些两亲物将DNA缩合成均匀的球形颗粒，其在还原时，膨胀成DNA环，随后释放游离DNA。复合物还原和DNA释放可以通过各种硫醇以及通过硫氧还蛋白还原酶酶促诱导。与DOTAP相比，在体外用两亲物-DNA复合物转染显示出转基因表达的数倍增加，并且可以通过将核靶向肽附接至两亲物来进一步增强。转染效率的增加是由于GSH和NAD（P）H依赖的复合物还原和释放游离DNA inside the cells.Conclusions：本工作证明了氧化还原控制的基因递送系统的原理，该系统使用硫醇-二硫键交换反应的可逆性。我们的数据表明，合成载体的效率可以通过它们在细胞内的受控不稳定来增强。由天然无毒化合物硫辛酸形成，这些阳离子两亲物为基因递送提供了一类新的有前途的合成载体。
Background: Intracellular release of free DNA from the vector complex is one of the critical steps limiting the efficiency of non-viral gene delivery. The complex should be stable enough to prevent DNA degradation but it should be destabilized inside the cell to allow DNA release and transcription. Destabilization and degradation of synthetic vectors is also required to reduce their cytotoxicity and augment the life-time of transfected cells.Results: Here we describe new cationic amphiphiles made from the natural provitamin, lipoic acid, that reversibly binds and releases DNA, depending on the redox state of the lipoate moieties, In the oxidized state these amphiphiles condense DNA into homogeneous spherical particles, which, upon reduction, swell into DNA toroids with subsequent release of free DNA. Complex reduction and DNA release can be induced by various thiols as well as enzymatically, by thioredoxin reductase. Transfection with amphiphile-DNA complexes in vitro shows a several fold increase of transgene expression compared with DOTAP, and can be further augmented by attachment of the nucleus-targeting peptide to the amphiphile. The increase of transfection efficiency results from GSH- and NAD(P)H-dependent complex reduction and release of free DNA inside the cells.Conclusions: The present work demonstrates the principle of a redox-controlled gene delivery system that uses the reversibility of thiol-disulfide exchange reaction. Our data suggest that the efficiency of synthetic vectors can be augmented by their controlled destabilization inside the cells. Being formed from the natural non-toxic compound lipoic acid, these cationic amphiphiles provide a new promising class of synthetic vectors for gene delivery.