Biointerphases

Journal for Biophysical Chemistry

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Open Access

Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing

  • Yee-Hung M. Chan1,
  • Bettina van Lengerich1 and
  • Steven G. Boxer1Email author
Biointerphases3:30200017

https://doi.org/10.1116/1.2889062

Received: 5 December 2007

Accepted: 7 February 2008

Abstract

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.