Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing
Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing
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DOI：
10.1116/1.2889062
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发表时间：
2008-06-01
期刊：
影响因子：
2.1
通讯作者：
Boxer, Steven G.
中科院分区：
文献类型：
--
作者：
Chan, Yee-Hung M.;van Lengerich, Bettina;Boxer, Steven G.
已经开发了用于合成5（'）-和3（'）-偶联的DNA-脂质缀合物的一般方法，并将其用于DNA介导的囊泡融合。呈现互补DNA的囊泡融合，导致内外小叶混合以及内容物混合。在相对的囊泡配偶体上使用5（'）-和3（'）-偶联的DNA而不是仅使用5（'）-偶联的DNA使融合最大化，这表明DNA取向对该过程的重要性。脂质和内容物混合测定显示每个囊泡的DNA序列和平均数的融合动力学的依赖性。没有DNA或呈现非互补序列的囊泡似乎也经历了一定程度的脂质混合或交换，但没有内容物混合。总脂质混合似乎比内瓣叶混合和内容物混合更有效，这可以通过观察到的非特异性脂质混合和/或半融合中间体的增加来解释。控制DNA序列的能力和该系统的相对实验简单性使得它非常有吸引力的探测膜融合使用合奏和单囊泡测定的基本问题。
A general method for synthesizing 5(')- and 3(')-coupled DNA-lipid conjugates has been developed and employed in DNA-mediated vesicle fusion. Vesicles presenting complementary DNA fuse, resulting in both outer and inner leaflet mixing as well as content mixing. Fusion is maximized using 5(')- and 3(')-coupled DNA on opposite vesicle partners, rather than only 5(')-coupled DNA, showing the importance of DNA orientation to the process. Lipid and content mixing assays show a dependence of fusion kinetics on the sequence and average number of DNA per vesicle. Vesicles without DNA or presenting noncomplementary sequences also appear to undergo some degree of lipid mixing or exchange, but no content mixing. Total lipid mixing appears to occur more efficiently than inner leaflet mixing and content mixing, and this may be explained by the observed nonspecific lipid mixing and/or the rise of a hemifused intermediate. The ability to control DNA sequence and the relative experimental simplicity of this system make it highly attractive to probe fundamental questions of membrane fusion using both ensemble and single vesicle assays.