Department of Biosciences & Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, 223-8522, Japan.
Chembiochem. 2021 Jul 1;22(13):2319-2324. doi: 10.1002/cbic.202100100. Epub 2021 May 10.
Giant unilamellar vesicles (GUVs) have been used as a material for bottom-up synthetic biology. However, due to the semi-permeability of the membrane, the need for methods to fuse GUVs has increased. To this aim, methods that are simple and show low leakage during fusion are important. In this study, we report a method of GUV fusion by a divalent cation (Ca ) enhanced with a long chain polyethylene glycol (PEG20k). The methods showed significant GUV fusion without leakage of internal components of GUVs and maintained cell-free transcription-translation ability inside the GUVs without external supplementation of macromolecules. We demonstrate that the Ca-PEG method can be applied for switching ON of transcription-translation in GUVs in a fusion-dependent manner. The method developed here can be applied to extend bottom-up synthetic biology and molecular robotics that use GUVs as a chassis.
巨大的单层囊泡 (GUVs) 已被用作从下而上的合成生物学的材料。然而，由于膜的半透性，对融合 GUVs 的方法的需求增加了。为此，简单且在融合过程中漏出率低的方法很重要。在这项研究中，我们报告了一种通过二价阳离子 (Ca 2+ ) 增强长链聚乙二醇 (PEG20k) 融合 GUV 的方法。该方法显示出明显的 GUV 融合，而没有 GUV 内部成分的泄漏，并在没有外部大分子补充的情况下保持 GUV 内的无细胞转录-翻译能力。我们证明 Ca-PEG 方法可以以依赖融合的方式用于 GUV 中转录-翻译的 ON 转换。这里开发的方法可用于扩展使用 GUVs 作为底盘的从下而上的合成生物学和分子机器人学。
