Ferrocene-containing cationic lipids for the delivery of DNA: Oxidation state determines transfection activity
Ferrocene-containing cationic lipids for the delivery of DNA: Oxidation state determines transfection activity
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DOI：
10.1016/j.jconrel.2006.01.014
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发表时间：
2006-05-01
影响因子：
10.8
通讯作者：
Lynn, DM
中科院分区：
文献类型：
--
作者：
Jewell, CM;Hays, ME;Lynn, DM
研究了两种具有氧化还原活性的含二茂铁阳离子脂质[11-（二茂铁十一烷基）三甲基溴化铵（FTMA）和双（11-二茂铁十一烷基）二甲基溴化铵（BFDMA）]转染哺乳动物细胞的能力。本研究试图确定这些脂质能够转染细胞的条件范围，以及这些材料中二茂铁基的氧化状态是否可以用来影响转染的程度。在COS-7细胞系中进行的实验表明，还原和氧化的FTMA基本上具有细胞毒性，并且不会转染细胞。随后使用BFDMA和编码增强型绿色荧光蛋白（EGFP）和萤火虫荧光素酶的报告质粒进行的实验表明，BFDMA能够转染细胞。然而，转染程度显著取决于BFDMA浓度和脂质氧化状态。细胞毒性和基因表达的定量表征表明，存在一个浓度窗口，超过这个浓度窗口，减少的BFDMA没有细胞毒性，产生高水平的转染，但超过这个浓度窗口，电化学氧化的BFDMA产生非常低（背景）的转染水平。利用动态光散射表征脂质团表明，在2 - 10 μ m的脂质浓度范围内，还原和氧化的BFDMA形成小聚集体（约90 - 250nrn）。综上所述，这些结果表明，BFDMA的氧化状态可以通过电化学控制，可以用来控制细胞转染的程度。这些结果可以形成转染程序的基础，利用含二茂铁脂质的氧化还原行为，利用电化学方法实现对转染的主动空间和时间控制。(c) 2006 Elsevier B.V.版权所有
The ability of two redox-active, ferrocene-containing cationic lipids [11-(ferrocenylundecyl)trimethylammonium bromide (FTMA) and bis(11-ferroceny lundecyl)dimethylammonium bromide (BFDMA)] to transfect mammalian cells was investigated. This study sought to determine the range of conditions over which these lipids were capable of transfecting cells and whether the oxidation state of the ferrocenyl groups in these materials could be used to influence the extent of transfection. Experiments conducted in the COS-7 cell line demonstrated that reduced and oxidized FTMA were substantially cytotoxic and did not transfect cells. Subsequent experiments conducted using BFDMA and reporter plasmids encoding enhanced green fluorescent protein (EGFP) and firefly luciferase demonstrated that BFDMA was able to transfect cells. However, the extent of transfection depended significantly upon both the concentration of BFDMA and the oxidation state of the lipid. Quantitative characterization of cytotoxicity and gene expression demonstrated that a window of concentration existed over which reduced BFDMA was noncytotoxic and yielded high levels of transfection, but over which electrochemically oxidized BFDMA yielded very low (background) levels of transfection. Characterization of lipoplexes using dynamic light scattering demonstrated that reduced and oxidized BFDMA formed small aggregates (ca. 90 to 250nrn) at concentrations of lipid ranging from 2 to 10 mu M. Taken together, these results demonstrate that the oxidation state of BFDMA, which can be controlled electrochemically, can be used to control the extent of cell transfection. These results could form the basis of transfection procedures that exploit the redox behavior of ferrocene-containing lipids to achieve active spatial and temporal control over transfection using electrochemical methods. (c) 2006 Elsevier B.V. All rights reserved.