Cerasome as an infusible and cell-friendly gene carrier: synthesis of cerasome-forming lipids and transfection using cerasome
Cerasome as an infusible and cell-friendly gene carrier: synthesis of cerasome-forming lipids and transfection using cerasome
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DOI：
10.1038/nprot.2006.182
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发表时间：
2006-10
期刊：
影响因子：
14.8
通讯作者：
Y. Sasaki;K. Matsui;Y. Aoyama;J. Kikuchi
中科院分区：
文献类型：
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作者：
Y. Sasaki;K. Matsui;Y. Aoyama;J. Kikuchi
超声处理在季铵化的氮上具有（EtO）3SiCH 2CH 2CH 2基团的阳离子脂质的预搅拌水溶液产生部分二氧化硅或陶瓷涂覆的脂质体（cerasome），其可用作优异的转染剂。非甲硅烷基化的参考脂质，其可以代表在常规脂质转染实验中使用的阳离子脂质，形成紧凑的脂质体，其经历DNA诱导的融合以提供与转染无关的和更大（100-300 nm）、毒性更强的颗粒。表面硬化的神经酰胺酶体是不熔的，并且DNA的单体神经酰胺酶体复合物具有病毒大小（约70 nm），并且表现出显著的转染性能，具有102-103倍的效率（相对于非甲硅烷基化的参比脂质）、最小化的细胞毒性和血清相容性。所述神经酰胺酶体脂质通过3-溴丙基三乙氧基硅烷与所述脂质的叔胺衍生物的反应获得。神经酰胺酶体水溶液的制备需要1-2小时。神经酰胺酶体-DNA复合物和转染大约需要3d完成。
Sonication of a pre-agitated aqueous solution of cationic lipid having a (EtO)3SiCH2CH2CH2group on the quarternized ammonium nitrogen results in partially silica- or ceramic-coated liposome (cerasome), which can be used as an excellent transfection agent. Non-silylated reference lipid, which may represent cationic lipids that are used in conventional lipofection experiments, form a compact liposome, which undergoes DNA-induced fusion to provide transfection-irrelevant and larger (100–300 nm), more toxic particles. The surface-rigidified cerasome is infusible and the monomeric cerasome complex of DNA is of viral size (∼70 nm) and exhibits a remarkable transfection performance with a 102–103-fold higher efficiency (relative to the non-silylated reference lipid), minimized cytotoxicity and serum compatibility. The cerasome lipid is obtained by the reaction of 3-bromopropyltriethoxysilane with a tertiary amine derivative of the lipid. Preparation of an aqueous cerasome solution takes 1–2 h. The cerasome–DNA complex and the transfection takes about 3 d to complete.