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Cerasome as an infusible and cell-friendly gene carrier: synthesis of cerasome-forming lipids and transfection using cerasome

Nature Protocols volume 1, pages 1227–1234 (2006)Cite this article

Abstract

Sonication of a pre-agitated aqueous solution of cationic lipid having a (EtO)3SiCH2CH2CH2 group 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.

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Figure 1
Figure 2: Preparation of the cerasome lipid.
Figure 3: Transmission electron microscopy imaging of single-walled cerasomes.
Figure 4: Size distribution profiles.
Figure 5: Electrophoretic gel shifts for pGL3 (160 ng).

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Acknowledgements

This work was supported by the Grant-in-Aid for Scientific Research (KAKENHI) in Priority Area “Molecular Nano Dynamics” (No. 17034026 for Y.A. and 17034043 for J.K.) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Graduate School of Materials Science, Nara Institute of Science and Technology (NAIST), 8916-5 Takayama, Ikoma, 630-0192, Nara, Japan

    Yoshihiro Sasaki & Jun-ichi Kikuchi

  2. Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan

    Kazuki Matsui & Yasuhiro Aoyama

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  1. Yoshihiro Sasaki
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Correspondence to Yasuhiro Aoyama or Jun-ichi Kikuchi.

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Sasaki, Y., Matsui, K., Aoyama, Y. et al. Cerasome as an infusible and cell-friendly gene carrier: synthesis of cerasome-forming lipids and transfection using cerasome. Nat Protoc 1, 1227–1234 (2006). https://doi.org/10.1038/nprot.2006.182

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