Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense, Denmark.
Methods Mol Biol. 2020;2105:75-96. doi: 10.1007/978-1-0716-0243-0_5.
Peptide nucleic acids (PNAs) can be modified with aliphatic lipid chains and designed to be water soluble and able to spontaneously insert into phospholipid bilayers. Liposomes with 1.5% negatively charged POPG can be driven to fuse and mix their inner content volumes via functionalization with such lipidated peptide nucleic acids (LiPNAs). During fusion, only low amounts of leakage occur (<5%). We describe here the synthesis and purification of such LiPNAs using an automated peptide synthesizer and the preparation of LiPNA functionalized liposomes. Further, we describe the measurement of LiPNA-induced fusion using a fluorescence-based assay for the content mixing between a liposome population with an encapsulated self-quenching fluorescent dye (SRB) and a buffer-filled liposome population.
肽核酸（PNA）可以用脂链修饰，并设计为水溶性，能够自发插入磷脂双层。带有 1.5%带负电荷的 POPG 的脂质体可以通过与这种脂化肽核酸（LiPNA）的功能化来驱动融合并混合它们的内部内容物体积。在融合过程中，只有少量泄漏发生（<5%）。在这里，我们描述了使用自动化肽合成仪合成和纯化这种 LiPNA 的方法，以及制备 LiPNA 功能化脂质体的方法。此外，我们还描述了使用荧光法测量 LiPNA 诱导的融合，该方法用于测量含有封装自猝灭荧光染料（SRB）的脂质体群体和充满缓冲液的脂质体群体之间的内容物混合。
